Use of lipid conjugates in the treatment of disease

ABSTRACT

This invention provides a method of preventing asthma, allergic rhinitis, or chronic obstructive pulmonary disease in a subject comprising the step of administering to a subject a compound comprising a lipid or phospholipid moiety bond to a physiologically acceptable monomer, dimer, oligomer, or polymer, and/or a pharmaceutically acceptable salt or a pharmaceutical product thereof. This invention also provides a method of treating allergic rhinitis or chronic obstructive pulmonary disease in a subject comprising the step of administering to a subject a compound comprising a lipid or phospholipid moiety bond to a physiologically acceptable monomer, dimer, oligomer, or polymer, and/or a pharmaceutically acceptable salt or a pharmaceutical product thereof.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part application of U.S.application Ser. No. 10/989,606, filed Nov. 17, 2004 now U.S. Pat. No.7,811,999 and of U.S. application Ser. No. 10/989,607, filed Nov. 17,2004, now U.S. Pat. No. 7,772,196 which are continuation-in-partapplications of U.S. application Ser. No. 10/627,981 filed Jul. 28,2003, now U.S. Pat. No. 7,101,859 which is a continuation-in-partapplication of U.S. application Ser. No. 09/756,765, filed Jan. 10,2001, now U.S. Pat. No. 7,034,006 which claims priority from aProvisional Application U.S. Ser. No. 60/174,907, filed Jan. 10, 2000;and a continuation-in-part application of PCT International ApplicationPCT/IL2005/001225, filed Nov. 17, 2005, which are hereby incorporated byreference.

FIELD OF THE INVENTION

This invention provides a method of preventing asthma, allergicrhinitis, or chronic obstructive pulmonary disease in a subjectcomprising the step of administering to a subject a compound comprisinga lipid or phospholipid moiety bond to a physiologically acceptablemonomer, dimer, oligomer, or polymer, and/or a pharmaceuticallyacceptable salt or a pharmaceutical product thereof. This invention alsoprovides a method of treating allergic rhinitis or chronic obstructivepulmonary disease in a subject comprising the step of administering to asubject a compound comprising a lipid or phospholipid moiety bond to aphysiologically acceptable monomer, dimer, oligomer, or polymer, and/ora pharmaceutically acceptable salt or a pharmaceutical product thereof.

BACKGROUND OF THE INVENTION

Lipid-conjugates having a pharmacological activity of inhibiting theenzyme phospholipase A2 (PLA2, EC 3.1.1.4) are known in the prior art.Phospholipase A2 catalyzes the breakdown of phospholipids at the sn-2position to produce a fatty acid and a lysophospholipid. The activity ofthis enzyme has been correlated with various cell functions,particularly with the production of lipid mediators such as eicosanoidproduction (prostaglandins, thromboxanes and leukotrienes), plateletactivating factor and lysophospholipids. Since their inception,lipid-conjugates have been subjected to intensive laboratoryinvestigation in order to obtain a wider scope of protection of cellsand organisms from injurious agents and pathogenic processes.

SUMMARY OF THE INVENTION

In one embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to prevent asthma.

In another embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to treat allergic rhinitis.

In another embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to prevent allergic        rhinitis.

In another embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to treat chronic        obstructive pulmonary disease,

In another embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to prevent chronic        obstructive pulmonary disease

In one embodiment, X in general formula (A) is a polysaccharide. In oneembodiment, the polysaccharide is carboxymethylcellulose, while inanother embodiment, the polysaccharide is a glycosaminoglycan. In oneembodiment, the glycosaminoglycan is hyaluronic acid, while in anotherembodiment, the glycosaminoglycan is heparin. In one embodiment L ingeneral formula (A) is phosphatidylethanolamine, which in one embodimentis dipalmitoyl phosphatidylethanolamine.

BRIEF DESCRIPTION OF FIGURES

FIG. 1.1: Inhibition of endothelin-1 (ET)-induced contraction of rattracheal rings by Lipid-conjugates. A: Contraction of rat trachea byEndothelin-1. B: Effect of HyPE on ET-induced contraction of rattrachea.

FIG. 1.2: Effect of HyPE and Hyaluronic acid (HA) on ET-1 inducedcontraction of rat trachea.

FIG. 1.3: Effect of HyPE and Hyaluronic acid (HA) on Acetylcholine(AcCh)-induced contraction of isolated rat trachea rings.

FIG. 1.4: Effect of HyPE, administered subcutaneously, on earlyasthmatic reaction (EAR) induced by ovalbumin inhalation

FIG. 1.5: Effect of HyPE on sPLA₂ expression in lung of rats withOVA-induced asthma.

FIG. 1.6: Effect of HyPE on cysteinyl leukotriens (LTC₄, LTD₄ and LTE₄)level in the BAL of OVA-induced asthmatic rats.

FIG. 1.7: Effect of HyPE inhalation on early and late asthmatic reaction(EAR and LAR, respectively) in OVA-sensitized asthmatic rats.

FIG. 1.8: Effect of HyPE inhalation on cysteinyl leukotriens (LTC4, LTD4and LTE4) level in the BAL of OVA-sensitized asthmatic rats.

FIG. 1.9: Effect of HyPE inhalation on NO production by macrophagescollected from the BAL of OVA-sensitized asthmatic rats.

FIG. 1.10: Effect of HyPE inhalation on structural change in airways(airway remodeling) of OVA sensitized asthmatic rats.

FIG. 1.11: Effect of HyPE on remodeling of asthmatic rat airway;histological morphometry.

FIG. 1.12: Effect of HyPE inhalation on TNFα production by macrophagescollected from the BAL of OVA-sensitized asthmatic rats.

FIG. 1.13: Amelioration of OVA-induced broncho-constriction by HyPEinhalation before challenge.

FIG. 14: Amelioration of OVA-induced broncho-constriction by HyPEinhalation after challenge.

FIG. 2.1: CMPE protects BGM cells from membrane lysis induced bycombined action of hydrogen peroxide (produced by glucose oxidase=GO),and exogenous phospholipase A₂ (PLA₂).

FIG. 2.2: CMPE protects BGM cells from glycosaminoglycan degradation byHydrogen peroxide (produced by GO).

FIG. 2.3: HyPE protects LDL from copper-induced oxidation.

FIG. 3.1: Effect of different Lipid-conjugates on LPS-induced IL-8production.

FIG. 3.2: Effect of HyPE on LPS-induced chemokine production.

FIG. 3.3: Effect of HyPE on LTA-induced IL-8 production.

FIG. 3.4: Effect of HyPE on LPS-induced ICAM-1 and E-selectinexpression.

FIG. 3.5: Effect of HyPE on LPS-induced activation of NF-kB in LMVEC.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides lipid-conjugates which display a wide-rangecombination of cytoprotective pharmacological activities. Thesecompounds can alleviate airway obstruction in asthma, protect mucosaltissue in gastrointestinal disease, suppress immune responses, alleviatecutaneous hypersensitivity reactions, inhibit cell proliferationassociated with vascular injury and immunological responses, inhibitcell migration associated with vascular and central nervous systemdisease, attenuate oxidative damage to tissue proteins and cellmembranes, interfere with viral spread, reduce tissue destroying enzymeactivity, and reduce intracellular levels of chemokines and cytokines.Thus these compounds are useful in the treatment of a diversity ofdisease states, including asthma, rhinitis, allergic rhinitis, chronicobstructive pulmonary disease, obstructive respiratory disease, colitis,Crohn's disease, central nervous system insult, multiple sclerosis,contact dermatitis, psoriasis, cardiovascular disease, invasive medicalprocedures, invasive cellular proliferative disorders, anti-oxidanttherapy, hemolytic syndromes, sepsis, acute respiratory distresssyndrome, tissue transplant rejection syndromes, autoimmune disease,viral infection, and hypersensitivity conjunctivitis

In another embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to prevent asthma.

In another embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid:    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to treat allergic rhinitis.

In another embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to prevent allergic        rhinitis.

In another embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to treat chronic        obstructive pulmonary disease.

In another embodiment, the invention provides for the use of a compoundrepresented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to prevent chronic        obstructive pulmonary disease.

In another embodiment, the present invention provides for the use of acompound represented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to treat a subject        suffering from an obstructive respiratory disease

In another embodiment, the present inventions provides for the use of acompound represented by the structure of the general formula (A):

-   -   wherein    -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000    -   for the preparation of a composition to prevent obstructive        respiratory disease.

In one embodiment, the obstructive respiratory disease is asthma. Inanother embodiment, the obstructive respiratory disease is rhinitis. Inanother embodiment, the obstructive respiratory disease is allergicrhinitis. In another embodiment, the obstructive respiratory disease ischronic obstructive pulmonary disorder. In another embodiment, theobstructive respiratory disease is sinusitis.

In one embodiment, X in general formula (A) is a polysaccharide. In oneembodiment, the polysaccharide is carboxymethylcellulose, while inanother embodiment, the polysaccharide is a glycosaminoglycan. In oneembodiment, the glycosaminoglycan is hyaluronic acid, while in anotherembodiment, the glycosaminoglycan is heparin. In one embodiment L ingeneral formula (A) is phosphatidylethanolamine, which in one embodimentis dipalmitoyl phosphatidylethanolamine.

In one embodiment, “treating” or “preventing” refers to delaying theonset of symptoms, reducing the severity of symptoms, reducing theseverity of an acute episode, reducing the number of symptoms, reducingthe incidence of disease-related symptoms, reducing the latency ofsymptoms, ameliorating symptoms, reducing secondary symptoms, reducingsecondary infections, prolonging patient survival, preventing relapse toa disease, decreasing the number or frequency of relapse episodes,increasing latency between symptomatic episodes, increasing time tosustained progression, expediting remission, inducing remission,augmenting remission, speeding recovery, or increasing efficacy of ordecreasing resistance to alternative therapeutics.

In one embodiment, symptoms are primary, while in another embodiment,symptoms are secondary. In one embodiment, “primary” refers to a symptomthat is a direct result of infection with a pathogen, while in oneembodiment, “secondary” refers to a symptom that is derived from orconsequent to a primary cause.

In one embodiment, the invention provides a method of treating a subjectsuffering from an obstructive respiratory disease, comprising the stepof administering to a subject a compound comprising a lipid orphospholipid moiety bond to a physiologically acceptable monomer, dimer,oligomer, or polymer, and/or a pharmaceutically acceptable salt or apharmaceutical product thereof, in an amount effective to treat thesubject suffering from an obstructive respiratory disease. In anotherembodiment, the invention provides a method of treating a subjectsuffering from an obstructive respiratory disease, comprising the stepof administering to a subject any one of the compounds according to theinvention, in an amount effective to treat the subject suffering from anobstructive respiratory disease. In another embodiment, the obstructiverespiratory disease is asthma.

In one embodiment of the invention, the physiologically acceptablemonomer is either a salicylate, salicylic acid, aspirin, amonosaccharide, lactobionic acid, maltose, an amino acid, glycine,carboxylic acid, acetic acid, butyric acid, dicarboxylic acid, glutaricacid, succinic acid, fatty acid, dodecanoic acid, didodecanoic acid,bile acid, cholic acid, cholesterylhemmisuccinate; or wherein thephysiologically acceptable dimer or oligomer is a dipeptide, adisaccharide, a trisaccharide, an oligopeptide, or a di- ortrisaccharide monomer unit of heparin, heparan sulfate, keratin, keratansulfate, chondroitin, chondoitin sulfate, dermatin, dermatan sulfate,dextran, or hyaluronic acid; or wherein the physiologically acceptablepolymer is a glycosaminoglycan, polygelin (‘hemaccell’), alginate,hydroxyethyl starch (hetastarch), polyethylene glycol, polycarboxylatedpolyethylene glycol, chondroitin sulfate, keratin, keratin sulfate,heparan sulfate, dermatin, dermatan sulfate, carboxymethylcellulose,heparin, dextran, or hyaluronic acid. In another embodiment, thephysiologically acceptable polymer is chondrotin sulfate. In anotherembodiment, the chondrotin sulfate is chondrotin-6-sulfate,chondroitin-4-sulfate or a derivative thereof. In another embodiment,the physiologically acceptable polymer is hyaluronic acid.

In one embodiment of the invention, the lipid or phospholipid moiety iseither phosphatidic acid, an acyl glycerol, monoacylglycerol,diacylglycerol, triacylglycerol, sphingosine, sphingomyelin,chondroitin-4-sulphate, chondroitin-6-sulphate, ceramide,phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine,phosphatidylinositol, or phosphatidylglycerol, or an ether or alkylphospholipid derivative thereof, and the physiologically acceptablemonomer or polymer moiety is either aspirin, lactobionic acid, maltose,glutaric acid, polyethylene glycol, carboxymethylcellulose, heparin,dextran, hemacell, hetastarch, or hyaluronic acid. In anotherembodiment, the phospholipid moiety is phosphatidylethanolamine

In one embodiment, obstructive respiratory disease is a disease ofluminal passages in the lungs, marked by dyspnea, tachypnea, orausculatory or radiological signs of airway obstruction. Obstructiverespiratory disease comprises asthma, acute pulmonary infections, acuterespiratory distress syndrome, chronic obstructive pulmonary disease,rhinitis, and allergic rhinitis. In one embodiment, the pathophysiologyis attributed to obstruction of air flow due to constriction of airwaylumen smooth muscle and accumulation of infiltrates in and around theairway lumen

In one embodiment, asthma is a disease process wherein the bronchi maybe narrowed, making breathing difficult. In one embodiment, symptomscomprise wheezing, difficulty breathing (particularly exhaling air),tightness in the chest, or a combination thereof. In one embodiment,factors which can exacerbate asthma include rapid changes in temperatureor humidity, allergies, upper respiratory infections, exercise, stress,smoke (cigarette), or a combination thereof.

In one embodiment, rhinitis comprises an inflammation of the mucousmembrane of the nose. In one embodiment, allergic rhinitis is aninflammatory response in the nasal passages to an allergic stimulus. Inone embodiment, symptoms comprise nasal congestion, sneezing, runny,itchy nose, or a combination thereof.

In one embodiment, chronic obstructive pulmonary disease is aprogressive disease process that most commonly results from smoking. Inone embodiment, chronic obstructive pulmonary disease comprisesdifficulty breathing, wheezing, coughing, which may be a chronic cough,or a combination thereof. In one embodiment, chronic obstructivepulmonary disease may lead to health complications, which in oneembodiment, may comprise bronchitis, pneumonia, lung cancer, or acombination thereof.

Colitis is a chronic disease of the gastrointestinal lumen, marked byabdominal discomfort, diarrhea and, upon radiological or histologicaldiagnosis, characteristic signs of mucosal damage including epithelialdenudation. Crohn's disease is a related disorder affecting typicallythe small intestine but which may involve any region of thegastrointestinal tract.

Multiple sclerosis is a disease of white matter, marked by motorweakness or sensory disturbance, or both, usually diagnosed by spinalfluid analysis or magnetic resonance imaging. Visual disturbance,including blindness, is common as well. In regions of disease activity,the blood brain barrier is impaired.

Skin hypersensitivity reactions, otherwise known as contact dermatitis,are marked by external signs of tissue irritation such as localizedredness, swelling, and pruritis. Virtually any substance may produce thecondition, and it is one of the most common complaints diagnosed bydermatologists.

Psoriasis is also one of the most common dermatologic diseases,affecting 1 to 2 percent of people. The most common areas of involvementare the elbows, knees, gluteal cleft, and the scalp. In active lesionsof psoriasis, the rate of epidermal cell replications is accelerated.Long-term use of topical glucocorticoids is often accompanied by loss ofeffectiveness,

Cardiovascular disease refers to both disorders of blood vessel lumennarrowing as well as to resultant ischemic syndromes of the targetorgans they supply, such as heart, kidney, and brain. Ischemia, orreduced blood supply, results from the narrowing of a blood vessel. Thesigns and symptoms of cardiovascular disease include, among others,angina pectoris, weakness, dyspnea, transient ischemic attacks, stroke,and renal insufficiency. Diagnosis is based on clinical grounds inconjunction with ancilliary diagnostic tests, such as blood tests,electrocardiograms, echography, and angiography. Atherosclerosis is acommon element in cardiovasular disease in which narrowing of the bloodvessel lumen is due to scar-like plaques formed from reactive,migrating, and proliferating cells and from local incorporation of bloodfat, cholesterol, and lipoprotein. Of particular significance in thisrespect is the accumulation of low density lipoprotein (LDL), which maybe accelerated when damaged by oxidation. Plaques are considered to bethe sites for both acute and chronic stenotic lesions, wherein the riskof tissue ischemia rises.

Stenotic or narrowing lesions of blood vessels occur not only inatherosclerosis but in other systemic cardiovascular disorders as well.Among these are arterial hypertension, vasculitides, including thevasculitis associated with transplanted organs, and coagulativedisorders. Many of these disorders, particularly hypertension,atherosclerosis, and vasculitis occur concommitantly in the samepatient.

Reperfusion injury and ischemia/reperfusion injury refers to the tissueinjury and initiation of necrosis following the resumption of blood flowto a previously ischemic tissue. This phenomenon is recognized as animportant component of ischemic and post-ischemic types of injury,particularly to brain and heart tissue. One pathophysiological mechanismwhich predominates in reperfusion is the damaging effect of reactiveoxygen species, otherwise known as oxidative damage or free radialinjury. Nitric oxide and its radicals are also implicated in thepathophysiology. The production of these noxious chemical species is isattributed to the local accumulation, adhesion, and transmigration ofleukocytes at the lesion site.

Invasive medical procedures, such as catheterization of arteries orveins or open surgery are frequently associated with tissue ischemia dueto blood vessel injury as well as to reperfusion injury, both of whichmay arise in the course of an invasive procedure. Thus preservation ofblood vessel potency and prevention of reperfusion injury are thesubject of intense investigation in medical science. Such procedures areperformed for both diagnostic and therapeutic purposes, and adjuvantdrugs are commonly prescribed to prevent complications of blood vesselinjury or restenosis. Formation of these lesions involves a multiplicityof participants, including coagulative elements of the blood, bloodcells, and the structural elements and cells of the blood vessel lumenwall. For example, arterial restenosis appearing after successfulballoon angioplasty is frequently due to the narrowing of the innerdiameter of the artery by the growth (proliferation) of smooth musclecells in the areas of irritation caused by the balloon angioplasty. Thisnew stenotic lesion may be comprised from other cell types as well,including leukocytes, accumulating at the lesion site through processesof migration and local proliferation. The two events (cell migration andproliferation) are almost certainly due to the coordinated interactionof a number of different cytokines likely released by early accumulationof macrophages at the site of original tissue injury. Thus leukocytescontribute to stenotic lesion formation through the processes ofmigration, local proliferation, passage through endothelial barriers,accumulation of cholesterol-rich lipoprotein, conversion to foam cells,and secretion of cytokines. This proliferation of cells and narrowing ofthe vascular lumen is not however restricted or limited to the coronaryarteries or cerebral circulation. It can also occur post-operativelycausing restenosis in, for example, peripheral vascular systems.

In the context of the present invention, the term cardiovascular diseaserefers to blood vessel lumen narrowing arising in the course ofatherosclerosis, vasculitis, invasive procedures, particularlycatheterization of an artery or vein, and the ischemic syndromesassociated with them.

Transplantation of tissue, grafts, and organs is frequently complicatedby the appearance of host-versus-graft and graft-versus-host disease,both of which may occur acutely or chronically in the recipient of thegraft. The source of the graft may be allogeneic (from the same species)or xenogeneic (from another species). Whether as complication due to theinduced hyperactive immune response, or through another mechanism,vasculitis remains a frequently encountered complication of tissuetransplantation procedures. Moreover, vascular damage due to reperfusioninjury is considered to be a major factor in the post-surgicalmalfunctioning of tissue and organ transplants.

Autoimmune diseases are conditions in which the change in clinical stateof the subject is attributed to aberrant cellular and/or humoral immuneresponses. The most common autoimmune diseases in the U.S. are juvenilediabetes, Hashimoto's and Grave's thryroiditis, rheumatoid arthritis,Crohn's disease and ulcerative colitis, chronic active hepatitis,vitaligo, glomerulonephritis, uveitis, multiple sclerosis, scleroderma,hemolytic anemia, idiopathic thrombocytopenic purpura, myastheniagravis, systemic lupus erythematosis, and pemphigus.

Hyper-proliferative cellular disorders, such as cancer cells arising atprimary organ sites or at other loci of spread (metastases), are one ofthe leading causes of death in the U.S. Cancers are frequently highlyresistant to all forms of treatment including therapy with potentanti-proliferative drugs and radiation. Increasingly the medicalcommunity is becoming aware of the critical role played by thevasculature associated with both the primary and metastatic forms ofdisease. Like any cell cluster, cancer cells are dependent upon areliable blood supply and in fact, cancer cells are known to encouragethe process of de novo vascularization through elaboration of growthfactors which act on endothelial cells and smooth muscle cells to formnew blood vessels, thus supplying the cancerous growth.

Metastasis, the spread of cancer cells to ectopic sites, is frequently avasculature dependent process as well, often referred to as hematogenousspread. The physiological barrier imposed by the blood vessel wall,comprised from elements such as endothelial cells and basement membranesubstance, is normally highly selective to the passage of cells.However, metastatic cells abrogate this barrier, employing a variety ofmechanisms, some of which have been established in the scientificliterature. For example, such abnormal cells produce hydrolytic enzymeswhich degrade the extracellular matrix and associated components of thevascular barrier, such as collagenase, heparinase, and hyaluronidase.Thus a critical factor in the metastatic process is the ability ofcancer cells to intrude through or permeate the wall of the blood vessellumen, thus arriving to invade a new tissue site after travel throughthe circulation. Cancer cells also elaborate messenger chemicals, knownas cytokines and chemokines, which enable the metastatic process, frommany aspects, including angiogenesis.

Cellular elaboration of cytokines and chemokines serve an importantregulatory function in health; however, when a hyperactive response tostress or disease is triggered, these compounds may present in excessand damage tissue, thereby pushing the disease state toward furtherdeterioration. Cytokine overproduction is involved in numerous diseases,such as sepsis, airway and lung injury, renal failure, transplantrejection, skin injuries, intestine injuries, cancer development andmetastasis, central nervous sytem disorders, vaginal bacterialinfection, and more. Two examples in which this occurs are systemicinfection, in particular when due to blood born bacteria (septicemia),and in the pulmonary condition known as acute (or adult) respiratorydistress syndrome (ARDS). In ARDS, lung spaces fill with fluid, impedinggas exchange and producing respiratory failure. Although plateletaggregation occurs, the major offenders appear to be monocyticphagocytes and leukocytes that adhere to endothelial surfaces andundergo a respiratory burst to inflict oxidant injury and releasechemokines such as Gro α, ENA-78, CX3X and MCP-1, in addition toleukotrienes, thromboxanes, and prostaglandins. The monocyticphagocytes, mainly macrophages in the alveoli and those lining thevasculature, also release oxidants, mediators, and a series ofdegradative enzymes that directly damage endothelial cells and causeleukocytes to release their lysosomal enzymes. The mortality rate isover 50%. The most common causes of ARDS are infection, aspiration,smoke and toxin inhalation, as well as systemic processes initiatedoutside the lung, including bacterial septicemia. The sepsis syndromeand shock are triggered by the interactions of various microbialproducts in the blood, in particular, gram-negative endotoxins, withhost mediator systems. The incidence is estimated to be up to 500,000cases per year in the U.S. alone, a figure which is considered to risedue to the increasing prevalence of antibiotic resistant organisms. Avariety of host mediators have been implicated in the pathogenesis ofsepticemia and septic shock (referred to collectively herein as sepsis)including factors released from stimulated cells, in particular,cytokines, tumor necrosis factor-α (TNF), Gro α, ENA-78, CX3X and MCP-1,NFκB transcription factor, lysosomal enzymes and oxidants fromleukocytes, and products of the metabolism of arachidonic acid, amongothers.

Red blood cell lysis, or hemolysis, may be an inherited or acquireddisorder, giving rise to anemia, iron deficiency, or jaundice. Among theacquired syndromes are membrane anomalies due to direct toxic effects ofsnake bites or of infectious agents, including viral, bacterial andparasitic etiologies, particularly malaria; exposure to oxidizingsubstances through ingestion or disease; or as a result of mechanicaltrauma within abnormal blood vessels. This latter condition, known asmicroangiopathic hemolysis, is considered to be related in mechanism tothe hemolysis produced from blood passage through prosthetic implants,such as heart valves. Inherited red blood cell membrane fragility oftenoccurs due to intracorpuscular enzyme and structural defects, such asglucose 6-phosphatase deficiency, sickle cell anemia, and thalessemia.Red blood cell lysis is one of the limiting factors in the storage lifeof blood products, particularly when subjected to free-radical formingphotodynamic virocidal treatments, such as γ-irradiation.

The acquired immunodeficiency syndrome is considered to be a rapidlygrowing global epidemic and one route of spread is through contaminatedblood products. Transmission and progression of this disease isdependent upon the infective activity of the human immunodeficiencyvirus. Current therapies are limited primarily to the administration ofreverse transcriptase inhibitors, drugs of high expense and low patienttolerability.

Oxidative injury refers to the effect of peroxidation and free radicalproduction on body tissues. To some extent, peroxide production is anormal, physiological process, attributed, for example, a role in immunedefense. However, in stress and disease states, or over the naturalcourse of time, as in physiological senesence, the accumulative additionof these unstable chemical moieties to tissue structures, includingmembrane components and blood proteins, leads to an irreversible patternof injury. Agents that act as anti-oxidants can protect againstoxidative damage. Such protection has been the subject of numerousscientific publications,

Intracellular bacterial parasites are one of the most prevelant forms ofsexually transmitted disease and are frequently intractable toconventional antibiotic therapy. Vaginal infection with chlamydiaspecies is a salient example.

In one embodiment, the present invention offers methods for thetreatment of disease based upon administration of lipids covalentlyconjugated through their polar head group to a physiologicallyacceptable chemical moiety, which may be of high or low molecularweight.

In one embodiment, the lipid compounds (Lipid-conjugates) of the presentinvention are described by the general formula:[phosphatidylethanolamine-Y]n-X[phosphatidylserine-Y]n-X[phosphatidylcholine-Y]n-X[phosphatidylinositol-Y]n-X[phosphatidylglycerol-Y]n-X[phosphatidic acid-Y]n-X[lyso-phospholipid-Y]n-X[diacyl-glycerol-Y]n-X[monoacyl-glycerol-Y]n-X[sphingomyelin-Y]n-X[sphingosine-Y]n-X[ceramide-Y]n-Xwherein

-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms; and-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer; and-   n, the number of lipid molecules bound to X, is a number from 1 to    1000.

In one embodiment of this invention, n is a number from 1 to 1000. Inanother embodiment, n is a number from 1 to 500. In another embodiment,n is a number from 1 to 100. In another embodiment, n is a number from100 to 300. In another embodiment, n is a number from 300 to 500. Inanother embodiment, n is a number from 500 to 800.

In one embodiment, the lipid compounds of this invention, known hereinas lipid conjugates (Lipid-conjugates) are now disclosed to possess acombination of multiple and potent pharmacological effects in additionto the ability to inhibit the extracellular form of the enzymephospholipase A2. The set of compounds comprisingphosphatidylethanolamine covalently bound to a physiologicallyacceptable monomer or polymer, is referred to herein as thePE-conjugates. Related derivatives, in which either phosphatidylserine,phosphatidylcholine, phosphatidylinositol, phosphatidic acid orphosphatidylglycerol are employed in lieu of phosphatidylethanolamine asthe lipid moiety provide equivalent therapeutic results, based upon thebiological experiments described below for the Lipid-conjugates and thestructural similarities shared by these compounds. Other Lipid-conjugatederivatives relevant to this invention are Lipid-conjugates wherein atleast one of the fatty acid groups of the lipid moieties at position C1or C2 of the glycerol backbone are substituted by a long chain alkylgroup attached in either ether or alkyl bonds, rather than esterlinkage.

As defined by the structural formulae provided herein for theLipid-conjugates, these compounds may contain between one to onethousand lipid moieties bound to a single physiologically acceptablepolymer molecule.

Administration of the Lipid-conjugates in a diversity of animal and cellmodels of disease invoices remarkable, and unexpected, cytoprotectiveeffects, which are useful in the treatment of disease. They are able tostabilize biological membranes; inhibit cell proliferation; suppressfree radical production; suppress nitric oxide production; reduce cellmigration across biological barriers; influence chemokine levels,including MCP-1, ENA-78, Gro α, and CX3C; affect gene transcription andmodify the expression of MHC antigens; bind directly to cell membranesand change the water structure at the cell surface; inhibit the uptakeof oxidized lipoprotein; prevent airway smooth muscle constriction;suppress neurotransmitter release; reduce expression of tumor necrosisfactor-α (TNF-α); modify expression of transcription factors such asNFκB; inhibit extracellular degradative enzymes, including collagenase,heparinase, hyaluronidase, in addition to that of PLA2; and inhibitviral infection of white cells. Thus the Lipid-conjugates providefar-reaching cytoprotective effects to an organism suffering from adisease wherein one or more of the presiding pathophysiologicalmechanisms of tissue damage entails either oxidation insult giving riseto membrane fragility; hyperproliferation behavior of cells giving riseto stenotic plaque formation in vascular tissue, angiogenesis and benignor malignant cancer disease, or psoriasis; aberrant cell migrationgiving rise to brain injury or tumor cell metastases; excessiveexpression of chemokines and cytokines associated with central nervoussystem (CNS) insult, sepsis, ARDS, or immunological disease; cellmembrane damage giving rise to CNS insult, CVS disease, or hemolysis;peroxidation of blood proteins and cell membranes giving rise toatherosclerosis or reperfusion injury; excessive nitric oxide productiongiving rise to CNS insult, reperfusion injury, and septic shock;interaction with major histocompatability antigens (MHC) associated withautoimmune diseases and alloimmune syndromes, such as transplantrejection.

In one embodiment of the present invention, the useful pharmacologicalproperties of the lipid or Lipid-conjugates may be applied for clinicaluse, and disclosed herein as methods for treatment of a disease. Thebiological basis of these methods may be readily demonstrated bystandard cellular and animal models of disease as described below.

While pharmacological activity of the Lipid-conjugates described hereinmay be due in part to the nature of the lipid moiety, the multiple anddiverse combination of pharmacological properties observed for theLipid-conjugates emerges ability of the compound structure to actessentially as several different drugs in one chemical entity. Thus, forexample, internal mucosal injury, as may occur in colitis or Crohn'sdisease, may be attenuated by any one or all of the pharmaceuticalactivities of immune suppression, anti-inflammation, anti-oxidation,nitric oxide production, or membrane stabilization. Protection of bloodvessels from periluminal damage, as may occur in atherosclerosis, mayentail activity from anti-proliferative, anti-chemokine, antioxidant, orantimigratory effects. Treatment or prevention of asthma, allergicrhinitis, chronic obstructive pulmonary disease, or obstructiverespiratory disease may involve any one of the many activities of theLipid-conjugates ranging from suppression of nitric oxide,anti-chemokine, anti-proliferative, or membrane stabilization effects.

Proliferation of vascular tissue is an element of both the atherogenesisof sclerotic plaques as well as a feature of primary and metastaticcancer lesion growth. Stabilization of biological membranes may preventhemolysis as well as mucosal bowel injury. Attenuation of chemokinelevels may ameliorate ADDS as well as militate against atherogenesis.Anti-oxidant activity protects may protect against reperfusion injuryand ischemia/reperfusion injury as well as CNS insult, atherosclerosis,and hemolysis. These and other advantages of the present invention willbe apparent to those skilled in the art based on the followingdescription.

The use of a single chemical entity with potent anti-oxidant,membrane-stabilizing, anti-proliferative, anti-chemokine,anti-migratory, and anti-inflammatory activity provides increasedcytoprotection relative to the use of several different agents each witha singular activity. The use of a single agent having multipleactivities over a combination or plurality of different agents providesuniform delivery of an active molecule, thereby simplifying issues ofdrug metabolism, toxicity and delivery. The compounds of the presentinvention also exhibit properties present only in the combined molecule,not in the individual components.

In one embodiment, the compounds of the invention may be used for acutetreatment of temporary conditions, or may be administered chronically,especially in the case of progressive, recurrent, or degenerativedisease. In one embodiment of the invention, the concentrations of thecompounds will depend on various factors, including the nature of thecondition to be treated, the condition of the patient, the route ofadministration and the individual tolerability of the compositions.

In another embodiment, the invention provides low-molecular weightLipid-conjugates, previously undisclosed and unknown to possesspharmacological activity, of the general formula:[Phosphatidylethanolamine-Y]n-X[Phosphatidylserine-Y]n-X[Phosphatidylcholine-Y]n-X[Phosphatidylinositol-Y]n-X[Phosphatidylglycerol-Y]n-X[Phosphatidic acid-Y]n-X[lyso-phospholipid-Y]n-X[diacyl-glycerol-Y]n-X[monoacyl-glycerol-Y]n-X[sphingomyelin-Y]n-X[sphingosine-Y]n-X[ceramide-Y]n-X

-   wherein-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms; and-   X is salicylate, salicylic acid, aspirin, a monosaccharide,    lactobionic acid, maltose, an amino acid, glycine, carboxylic acid,    acetic acid, butyric acid, dicarboxylic acid, glutaric acid,    succinic acid, fatty acid, dodecanoic acid, didodecanoic acid, bile    acid, cholic acid, cholesterylhemmisuccinate, a dipeptide, a    disaccharide, a trisaccharide, an oligosaccharide, an oligopeptide,    or a di- or trisaccharide monomer unit of heparin, heparan sulfate,    keratin, keratan sulfate, chondroitin, chondoitin-6-sulfate,    chondroitin-4-sulfate, dermatin, dermatan sulfate, dextran, or    hyaluronic acid, a glycosaminoglycan, polygeline (‘haemaccel’),    alginate, hydroxyethyl starch (hetastarch), polyethylene glycol,    polycarboxylated polyethylene glycol, chondroitin-6-sulfate,    chondroitin-4-sulfate, keratin, keratin sulfate, heparan sulfate,    dermatin, dermatan sulfate, carboxymethylcellulose, heparin,    dextran, or hyaluronic acid; and n, the number of lipid molecules    bound to X, is a number from 1 to 1000

In one embodiment of this invention, n is a number from 1 to 1000. Inanother embodiment, n is a number from 1 to 500. In another embodiment,n is a number from 1 to 100. In another embodiment, n is a number from100 to 300. In another embodiment, n is a number from 300 to 500. Inanother embodiment, n is a number from 500 to 800.

In another embodiment of the invention, these Lipid-conjugatederivatives possess wide-spectrum pharmacological activity and, aspharmaceutical agents administered to treat disease, are consideredanalogous to the Lipid-conjugates comprised from high molecular weightpolymers. Other lipid-conjugate derivatives relevant to this inventionare glycerolipid moieties in which at least one of the two long chainalkyl groups in position C1 and C2 of the glycerol backbone are attachedin ether or alkyl bonds, rather than ester linkage.

The present invention is further illustrated in the following examplesof the therapeutic Lipid-conjugate compounds, their chemicalpreparation, their anti-disease activity, and methods of use aspharmaceutical compositions in the treatment of disease

Compounds

In the methods, according to embodiments of the invention, theLipid-conjugates administered to the subject are comprised from at leastone lipid moiety covalently bound through an atom of the polar headgroup to a monomer or polymeric moiety (referred to herein as theconjugated moiety) of either low or high molecular weight. When desired,an optional bridging moiety can be used to link the Lipid-conjugatesmoiety to the monomer or polymeric moiety. The conjugated moiety may bea low molecular weight carboxylic acid, dicarboxylic acid, fatty acid,dicarboxylic fatty acid, acetyl salicylic acid, cholic acid,cholesterylhemisuccinate, or mono- or di-saccharide, an amino acid ordipeptide, an oligopeptide, a glycoprotein mixture, a di- ortrisaccharide monomer unit of a glycosaminoglycan such as a repeatingunit of heparin, heparan sulfate, hyaluronic acid, chondrotin-sulfate,dermatan, keratan sulfate, or a higher molecular weight peptide oroligopeptide, a polysaccharide, polyglycan, protein, glycosaminoglycan,or a glycoprotein mixture. From a composition aspect,phospholipid-conjugates of high molecular weight, and associatedanalogues, are the subject of U.S. Pat. No. 5,064,817, as well as thepublications cited herein.

In one embodiment of the invention, when the conjugated carrier moietyis a polymer, the ratio of lipid moieties covalently bound may rangefrom one to one thousand lipid residues per polymer molecule, dependingupon the nature of the polymer and the reaction conditions employed. Forexample, the relative quantities of the starting materials, or theextent of the reaction time, may be modified in order to obtainLipid-conjugate products with either high or low ratios of lipidresidues per polymer, as desired.

The term “moiety” means a chemical entity otherwise corresponding to achemical compound, which has a valence satisfied by a covalent bond.

Examples of polymers which can be employed as the conjugated moiety forproducing Lipid-conjugates for use in the methods of this invention maybe physiologically acceptable polymers, including water-dispersible or-soluble polymers of various molecular weights and diverse chemicaltypes, mainly natural and synthestic polymers, such asglycosaminoglycans, hyaluronic acid, heparin, heparin sulfate,chondrotin sulfate, chondrotin-6-sulfate, chondroitin-4-sulfate, keratinsulfate, dermatin, sulfate, plasma expanders, including polygeline(“Haemaccel”, degraded gelatin polypeptide crosslinked via urea bridges,produced by “Behring”), “hydroxyethylstarch” (Htastarch, HES) andextrans, food and drug additives, soluble cellulose derivatives (e.g.,methylcellulose, carboxymethylcellulose), polyaminoacids, hydrocarbonpolymers (e.g., polyethylene), polystyrenes, polyesters, polyamides,polyethylene oxides (e.g. polyethyleneglycols;polycarboxyethyleneglycol), polyvinnylpyrrolidones, polysaccharides,alginates, assimilable gums (e.g., xanthan gum), peptides, injectableblood proteins (e.g., serum albumin), cyclodextrin, and derivativesthereof.

Examples of monomers, dimers, and oligomers which can be employed as theconjugated moiety for producing Lipid-conjugates for use in the methodsof the invention may be mono- or disaccharides, carboxylic acid,dicarboxylic acid, fatty acid, dicarboxylic fatty acid, acetyl salicylicacid, cholic acid, cholesterylhemisuccinate, and di- and trisaccharideunit monomers of glycosaminoglycans including heparin, heparan sulfate,hyaluronic acid, chondrotin, chondioitin-6-sulfate,chondroitin-4-sulfate, dermatin, dermatan sulfate, keratin, keratansulfate, or dextran.

In some cases, according to embodiments of the invention, the monomer orpolymer chosen for preparation of the Lipid-conjugate may in itself haveselect biological properties. For example, both heparin and hyaluronicacid are materials with known physiological functions. In the presentinvention, however, the Lipid-conjugates formed from these substances asstarting materials display a new and wider set of pharmaceuticalactivities than would be predicted from administration of either heparinor hyaluronic acid which have not been bound by covalent linkage to aphospholipid. It can be shown, by standard comparative experiments asdescribed below, that phosphatidylethanolamine (PE) linked tocarboxymethylcellulose (referred to as CMPE, CMC-Peor CME), tohyaluronic acid (referred to as HYPE, HyPE, and Hyal-PE), to heparin(referred to as HEPPE, HepPE, HePPE, Hepa-PE), to chondroitine sulfate A(referred to as CSAPE, CsaPE, CsAPE), to Polygeline (haemaccel)(referred to HemPE, HEMPE), or to hydroxyethyl starch (referred to asHesPE, HESPE), are far superior in terms of potency and range of usefulpharmaceutical activity to the free conjugates (the polymers above andthe like). In fact, these latter substances are, in general, notconsidered useful in methods for treatment of most of the diseasesdescribed herein, and for those particular cases wherein their use ismedically prescribed, such as ischemic vascular disease, theconcentrations for their use as drugs are are several orders ofmagnitude higher. Thus, the combination of a phospholipid such asphosphatidylethanolamine, or related phospholipids which differ withregard to the polar head group, such as phosphatidylserine (PS),phosphatidylcholine (PC), phosphatidylinositol (PI), andphosphatidylglycerol (PG), results in the formation of a compound whichhas novel pharmacological properties when compared to the startingmaterials alone

The biologically active lipid conjugates described herein can have awide range of molecular weight, e.g., above 50,000 (up to a few hundredthousands) when it is desirable to retain the Lipid conjugate in thevascular system and below 50,000 when targeting to extravascular systemsis desirable. The sole limitation on the molecular weight and thechemical structure of the conjugated moiety is that it does not resultin a Lipid-conjugate devoid of the desired biological activity, or leadto chemical or physiological instability to the extent that theLipid-conjugate is rendered useless as a drug in the method of usedescribed herein,

In one embodiment, the compound according to the invention isrepresented by the structure of the general formula (A):

wherein

-   L is a lipid or a phospholipid;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein X is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between L, Z, Y and X is either an amide or an    esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (I):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms; and    -   X is either a physiologically acceptable monomer, dimer,        oligomer or a physiologically acceptable polymer, wherein X is a        glycosaminoglycan; and    -   n is a number from 1 to 1,000;    -   wherein if Y is nothing the phosphatidylethanolamine is directly        linked to X via an amide bond and if Y is a spacer, the spacer        is directly linked to X via an amide or an esteric bond and to        the phosphatidylethanolamine via an amide bond.

Preferred compounds for use in the methods of the invention comprise oneof the following as the conjugated moiety X: acetate, butyrate,glutarate, succinate, dodecanoate, didodecanoate, maltose, lactobionicacid, dextran, alginate, aspirin, cholate, cholesterylhemisuccinate,carboxymethyl-cellulose, heparin, hyaluronic acid, polygeline(haemaccel), polyethyleneglycol, and polycarboxylated polyethyleneglycol. The polymers used as starting material to prepare thePE-conjugates may vary in molecular weight from 1 to 2,000 kDa.

Examples of phosphatidylethanolamine (PE) moieties are analogues of thephospholipid in which the chain length of the two fatty acid groupsattached to the glycerol backbone of the phospholipid varies from 2-30carbon atoms length, and in which these fatty acids chains containsaturated and/or unsaturated carbon atoms. In lieu of fatty acid chains,alkyl chains attached directly or via an ether linkage to the glycerolbackbone of the phospholipid are included as analogues of PE. Accordingto the present invention, a most preferred PE moiety isdipalmitoylphosphatidy-ethanolamine.

Phosphatidyl-ethanolamine and its analogues may be from various sources,including natural, synthetic, and semisynthetic derivatives and theirisomers.

Phospholipids which can be employed in lieu of the PE moiety areN-methyl-PE derivatives and their analogues, linked through the aminogroup of the N-methyl-PE by a covalent bond; N,N-dimethyl-PE derivativesand their analogues linked through the amino group of theN,N-dimethyl-PE by a covalent bond, phosphatidylserine (PS) and itsanalogues, such as palmitoyl-stearoyl-PS, natural PS from varioussources, semisynthetic PSs, synthetic, natural and artifactual PSs andtheir isomers. Other phospholipids useful as conjugated moieties in thisinvention are phosphatidylcholine (PC), phosphatidylinositol (PI),phosphatidic acid and phosphoatidylglycerol (PG), as well as derivativesthereof comprising either phospholipids, lysophospholipids,phosphatidyic acid, sphingomyel ins, lysosphingomyel ins, ceramide, andsphingosine.

For PE-conjugates and PS-conjugates, the phospholipid is linked to theconjugated monomer or polymer moiety through the nitrogen atom of thephospholipid polar head group, either directly or via a spacer group.For PC, PI, and PG conjugates, the phospholipid is linked to theconjugated monomer or polymer moiety through either the nitrogen or oneof the oxygen atoms of the polar head group, either directly or via aspacer group.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (II):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein if Y is nothing the phosphatidylserine is directly linked to    X via an amide bond and if Y is a spacer, the spacer is directly    linked to X via an amide or an esteric bond and to the    phosphatidylserine via an amide bond.    In another embodiment, the compound according to the invention be    [phosphatidylserine-Y]n-X, wherein Y is either nothing or a spacer    group ranging in length from 2 to 30 atoms, X is a physiologically    acceptable monomer, dimer, oligomer or polymer wherein x is a    glycosaminoglycan, and n is a number from 1 to 1000, wherein the    phosphatidylserine may be bonded to Y or to X, if Y is nothing, via    the COO⁻ moiety of the phosphatidylserine.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (III):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phosphatidyl, Z, Y and X is either an    amide or anesteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (IV):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (V):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (VI):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either, nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (VII):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond.

In one embodiment of the invention, phosphatidylcholine (PC),Phosphatidylinositol (PI), phosphatidic acid (PA), wherein Z is nothing,and Phosphatidylglycerol (PG) conjugates are herein defined as compoundsof the general formula (III).

In one embodiment of the invention Y is nothing Non limiting examples ofsuitable divalent groups forming the optional bridging group (spacer) Y,according to embodiments of the invention, are straight or branchedchain alkylene, e.g., of 2 or more, preferably 4 to 30 carbon atoms,—CO-alkylene-CO, —NH-alkylene-NH—, —CO-alkylene-NH—,—NH-alkylene-NHCO-alkylene-NH—, an amino acid, cycloalkylene, whereinalkylene in each instance, is straight or branched chain and contains 2or more, preferably 2 to 30 atoms in the chain, —(—O—CH(CH₃)CH₂—)_(x)—wherein x is an integer of 1 or more.

According to embodiments of the invention, in addition to thetraditional phospholipid structure, related derivatives for use in thisinvention are phospholipids modified at the C1 or C2 position to containan ether or alkyl bond instead of an ester bond. In one embodiment ofthe invention, the alkyl phospholipid derivatives and ether phospholipidderivatives are exemplified herein.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (VIII):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (IX):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (IXa):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bonds

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (IXb):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (X):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimes, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the ceramide phosphoryl, Z, Y and X is    either an amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XI):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein if Y is nothing the sphingosyl is directly linked to X via    an amide bond and if Y is a spacer, the spacer is directly linked to    X and to the sphingosyl via an amide bond and to X via an amide or    an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XII):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   L is ceramide;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the ceramide, Z, Y and X is either an amide    or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XIII):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the diglyceryl, Z, Y and X is either an    amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XIV):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the glycerolipid, Z, Y and X is either an    amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XV):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the glycerolipid, Z, Y and X is either an    amide or an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XVI):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XVII):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XVIII):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XIX):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer; oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XX):

wherein

-   R₁ is either hydrogen or a linear-saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond.

In another embodiment, the compound according to the invention isrepresented by the structure of the general formula (XXI):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond

In one embodiment of the invention, the glycosaminoglycan may be, interalia, hyaluronic acid, heparin, heparan sulfate, chondrotin sulfate,keratin, keratan sulfate, dermatan sulfate or a derivative thereof.

In another embodiment, the glycosaminoglycan is di- and trisaccharideunit monomers of glycosaminoglycans. In another embodiment, thechondroitin sulfate may be, inter alia, chondroitin-6-sulfate,chondroitin-4-sulfate or a derivative thereof.

In one embodiment of the invention, the sugar rings of theglycosaminoglycan are intact. In another embodiment, intact refers toclosed. In another embodiment, intact refers to natural. In anotherembodiment, intact refers to unbroken.

In one embodiment of the invention, the structure of the lipid orphospholipids in any compound according to the invention is intact. Inanother embodiment, the natural structure of the lipid or phospholipidsin any compound according to the invention is maintained.

In one embodiment, the compounds according to the invention arebiodegradable.

In one embodiment, the compound according to the invention is a compoundrepresented by the structure of the general formula (A):

wherein

-   L is phosphatidyl;-   Z is ethanolamine, wherein L and Z are chemically bonded resulting    in phosphatidylethanolamine;-   Y is nothing;-   X is hyaluronic acid; and-   n is a number from 1 to 1000;    wherein any bond between the phosphatidylethanolamine and the    hyaluronic acid is an amide bond.

In one embodiment, the compound according to the invention is a compoundrepresented by the structure of the general formula (A):

wherein

-   L is phosphatidyl;-   Z is ethanolamine, wherein L and Z are chemically bonded resulting    in phosphatidylethanolamine;-   Y is nothing;-   X is chondroitin sulfate; and-   n is a number from 1 to 1000;    wherein any bond between the phosphatidylethanolamine and the    chondroitin sulfate is an amide bond.

In another embodiment, the invention provides a method of treating asubject suffering from asthma, comprising the step of administering to asubject any one of the compounds according to the invention, or anycombination thereof, in an amount effective to treat the subjectsuffering from asthma. In another embodiment, the compounds according tothe invention include, inter alia, the compounds represented by thestructures of the general formulae: (A), (I), (II), (III), (IV), (V),(VI), (VII), (VIII), (IX), (IXa), (IXb), (X), (XI), (XII), (XIII),(XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX), (XXI), (XXII) or anycombination thereof. In another embodiment, the invention provides amethod of preventing asthma in a subject.

In another embodiment, the invention provides a method of treating asubject suffering from rhinitis, comprising the step of administering toa subject any one of the compounds according to the invention, or anycombination thereof, in an amount effective to treat the subjectsuffering from rhinitis. In another embodiment, the compounds accordingto the invention include, inter alia, the compounds represented by thestructures of the general formulae: (A), (I), (II), (III), (IV), (V),(VI), (VII), (VIII), (IX), (IXa), (IXb), (X), (XI), (XII), (XIII),(XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX), (XXI), (XXII) or anycombination thereof. In another embodiment, the invention provides amethod of preventing rhinitis in a subject.

In another embodiment, the invention provides a method of treating asubject suffering from allergic rhinitis, comprising the step ofadministering to a subject any one of the compounds according to theinvention, or any combination thereof, in an amount effective to treatthe subject suffering from allergic rhinitis. In another embodiment, thecompounds according to the invention include, inter alia, the compoundsrepresented by the structures of the general formulae: (A), (I), (II),(III), (IV), (V), (VI), (VII), (VIII), (IX), (IXa), (IXb), (X), (XI),(XII), (XIII), (XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX), (XXI),(XXII) or any combination thereof. In another embodiment, the inventionprovides a method of preventing allergic rhinitis in a subject.

In another embodiment, the invention provides a method of treating asubject suffering from chronic obstructive pulmonary disease, comprisingthe step of administering to a subject any one of the compoundsaccording to the invention, or any combination thereof, in an amounteffective to treat the subject suffering from chronic obstructivepulmonary disease. In another embodiment, the compounds according to theinvention include, inter alia, the compounds represented by thestructures of the general formulae: (A), (I), (II), (III), (IV), (V),(VI), (VII), (VIII), (IX), (IXa), (IXb), (X), (XI), (XII), (XIII),(XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX), (XXI), (XXII) or anycombination thereof. In another embodiment, the invention provides amethod of preventing chronic obstructive pulmonary disease in a subject.

In another embodiment, the invention provides a method of treating asubject suffering from an obstructive respiratory disease, comprisingthe step of administering to a subject any one of the compoundsaccording to the invention, or any combination thereof, in an amounteffective to treat the subject suffering from an obstructive respiratorydisease. In another embodiment, the compounds according to the inventioninclude, inter alia, the compounds represented by the structures of thegeneral formulae: (A), (I), (II), (III), (IV), (V), (VI), (VII), (VIII),(IX), (IXa), (IXb), (X), (XI), (XII), (XIII), (XIV), (XV), (XVI),(XVII), (XVIII), (XIX), (XX), (XXI), (XXII) or any combination thereof.In another embodiment, the obstructive respiratory disease is asthma. Inanother embodiment, the obstructive respiratory disease is rhinitis. Inanother embodiment, the obstructive respiratory disease is allergicrhinitis. In another embodiment, the obstructive respiratory disease ischronic obstructive pulmonary disease. In another embodiment, theinvention provides a method of preventing asthma, rhinitis, allergicrhinitis, chronic obstructive pulmonary disease, obstructive respiratorydisease or a combination thereof, in a subject.

Illustrative of preferred Lipid-conjugates for use in the methodsaccording to embodiments of this invention are those in which thelipid/phospholipid moiety is linked directly or indirectly through abridging moiety listed below.

phospholipid spacer polymer (m.w.) abbreviation PE DicarboxylicPolygeline HeMPE; HemPE acid + Diamine (haemaccel) (4-40 kDa) PE NoneCarboxymethyl- CMPE; CMC- cellulose PE (20-500 kDa) PE None HyaluronicHYPE (HyPE) acid (2-2000 kDa) PE Dipalmitoic Hyaluronic acid HYPE- acid(2-2000 kDa) dipalmitoyl PE None Polyethylene glycol PE Y Hydroxyethyl-HESPE; HesPE starch PE Dicarboxylic Dextran DexPE acid + Diamine(1-2,000 kDa) PE None Dextran DexPE (1-2,000 kDa) PE None Albumin PENone Alginate (2-2000 kDa) PE None Polyarninoacid PE None Lactobionicacid PE None Acetylsali- cylate PE None Cholesteryl- hemmisuccinate PENone Maltose PE Y None Cholic acid PE None Polycarboxylated polyethyleneglycol PE None Heparin HEPPE ;HEPE; (05-110 kDa) HepPE Dimyristoyl-PE YVariable DMPE Dimyristoyl-PE Y Hyaluronic acid HyDMPE PS Y Polygeline(haemaccel) PS Y Heparin PS Y Hyaluronic acid PC Y Polygeline(haemaccel) PC Y Hepariri PC Y Hyaluronic acid PI Y Polygeline(haemaccel) PI Y Heparin PI Y Hyaluronic acid PG Y Polygeline(haemaccel) PG Y Heparin PE Y Chondoitin sulfates CSPE PE Y Polygeline(haemaccel) PG Y Hyaluronic acid

In one embodiment of the invention, the compounds administered are HyPE,CSAPE, CMPE, HemPE, HesPE, DexPE and As-PE and pharmaceuticallyacceptable salts thereof, in combination with a physiologicallyacceptable carrier or solvent. These polymers, when chosen as theconjugated moiety, may vary in molecular weights from 200 to 2,000,000Daltons. Various molecular weight species have been shown to have thedesired biological efficacy, as shown in the section below.

In addition to the compounds of the Examples, further illustrativecompounds of this invention are set forth in the section below.

Novel Compounds

Low molecular weight Lipid-conjugates, in which the conjugated moiety isa monomer such as a salicylate, a bile acid, orcholesterylhemmisuccinate, or a di- or trisaccaharide unit monomer of apolyglycosoaminoglycan such as heparin, heparan sulfate,chondrotin-6-sulfate, chondroitin-4-sulfate, hyaluronic acid, keratin,keratan sulfate, dermatin, or dermatin sulfate, have not been describedbefore. According to embodiments of the invention, these new compoundsdisplay a similar biological activity profile as demonstrated below forthe other Lipid-conjugates and have the general formula[Phosphatidylethanolamine-Y]_(n)—X[Phosphatidylserine-Y]_(n)—X[Phosphatidylcholine-Y]_(n)—X[Phosphatidylinositol-Y]_(n)—X[Phosphatidylglycerol-Y]_(n)—X[Phosphatidic acid-Y]_(n)—X[lyso-phospholipid-Y]_(n)—X[diacyl-glycerol-Y]_(n)—X[monoacyl-glycerol-Y]_(n)—X[sphingomyelin-Y]_(n)—X[sphingosine-Y]_(n)—X[ceramide-Y]_(n)—Xwherein

-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a mono- or disaccharide, carboxylated disaccharide, mono- or    dicarboxylic acids, a salicylate, salicylic acid, aspirin,    lactobionic acid, maltose, an amino acid, glycine, acetic acid,    butyric acid, dicarboxylic acid, glutaric acid, succinic acid, fatty    acid, dodecanoic acid, didodecanoic acid, bile acid, cholic acid,    cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, a    trisacharide, or a di- or trisaccharide monomer unit of heparin,    heparan sulfate, keratin, keratan sulfate, chondroitin,    chondoitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatan    sulfate, dextran, or hyaluronic acid; and-   n is the number of lipid moiety molecules bound to a molecule of X    wherein n is a number from 1 to 1000.

In one embodiment of this invention, low molecular weightphosphatidylethanolamine (PE)-conjugates are defined hereinabove as thecompounds of formula (I) wherein:

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a mono- or disaccharide, carboxylated disaccharide, mono- or    dicarboxylic acids, a salicylate, salicylic acid, aspirin,    lactobionic acid, maltose, an amino acid, glycine, acetic acid,    butyric acid, dicarboxylic acid, glutaric acid, succinic acid, fatty    acid, dodecanoic acid, didodecanoic acid, bile acid, cholic acid,    cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, a    trisacharide, or a di- or trisaccharide monomer unit of heparin,    heparan sulfate, keratin, keratan sulfate, chondroitin,    chondoitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatan    sulfate, dextran, or hyaluronic acid; and-   n is the number of lipid moity molecules bound to a molecule of X    wherein n is a number from 1 to 1000.

In one embodiment of this invention, low molecular weightphosphatidylserine (PS)-conjugates are defined hereinabove as thecompounds of formula (II) wherein:

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a mono- or disaccharide, carboxylated disaccharide, mono- or    dicarboxylic acids, a salicylate, salicylic acid, aspirin,    lactobionic acid, maltose, an amino acid, glycine, acetic acid,    butyric acid, dicarboxylic acid, glutaric acid, succinic acid, fatty    acid, dodecanoic acid, didodecanoic acid, bile acid, cholic acid,    cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, a    trisaccharide, or a di- or trisaccharide monomer unit of heparin,    heparan sulfate, keratin, keratan sulfate, chondroitin,    chondoitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatan    sulfate, dextran, or hyaluronic acid; and-   n is the number of lipid moiety molecules bound to a molecule of X    wherein n is a number from 1 to 1000.

In one embodiment of this invention, Phosphatidylcholine (PC),Phosphatidylinositol (PI), and Phosphatidylglycerol (PG) conjugates arehereinabove defined as the compounds of formula (III) wherein:

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a mono- or disaccharide, carboxylated disaccharide, mono- or    dicarboxylic acids, a salicylate, salicylic acid, aspirin,    lactobionic acid, maltose, an amino acid, glycine, acetic acid,    butyric acid, dicarboxylic acid, glutaric acid, succinic acid, fatty    acid, dodecanoic acid, didodecanoic acid, bile acid, cholic acid,    cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, a    trisaccharide, or a di- or trisaccharide monomer unit of heparin,    heparan sulfate, keratin, keratan sulfate, chondroitin,    chondoitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatan    sulfate, dextran, or hyaluronic acid; and-   n is the number of lipid moiety molecules bound to a molecule of X    wherein n is a number from 1 to 1000.

Examples of suitable divalent groups forming the optional bridging groupY are straight- or branched-chain alkylene, e.g., of 2 or more,preferably 4 to 18 carbon atoms, —CO-alkylene-CO, —NH-alkylene-NH—,—CO-alkylene-NH—, cycloalkylene, wherein alkylene in each instance, isstraight or branched chain and contains 2 or more, preferably 2 to 18carbon atoms in the chain, —(—O—CH(CH₃)CH₂—)_(x)— wherein x is aninteger of 1 or more.

In another embodiment, in addition to the traditional phospholipidstructure, related derivatives for use in this invention arephospholipids modified at the C1 or C2 position to contain an ether oralkyl bond instead of an ester bond. These derivatives are exemplifiedhereinabove by the general formulae (VIII) and (IX) wherein:

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a mono- or disaccharide, carboxylated disaccharide, mono- or    dicarboxylic acids, a salicylate, salicylic acid, aspirin,    lactobionic acid, maltose, an amino acid, glycine, acetic acid,    butyric acid, dicarboxylic acid, glutaric acid, succinic acid, fatty    acid, dodecanoic acid, didodecanoic acid, bile acid, cholic acid,    cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, a    trisaccharide, or a di- or trisaccharide monomer unit of heparin,    heparan sulfate, keratin, keratan sulfate, chondroitin,    chondoitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatan    sulfate, dextran, or hyaluronic acid; and-   n is the number of lipid moity molecules bound to a molecule of X    wherein n is a number from 1 to 1000.

In another embodiment, related low molecular weight derivatives for usein this invention are exemplified hereinabove by the general formulae(X), (XI) and (XII) wherein:

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a mono- or disaccharide, carboxylated disaccharide, mono- or    dicarboxylic acids, a salicylate, salicylic acid, aspirin,    lactobionic acid, maltose, an amino acid, glycine, acetic acid,    butyric acid, dicarboxylic acid, glutaric acid, succinic acid, fatty    acid, dodecanoic acid, didodecanoic acid, bile acid, cholic acid,    cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, a    trisaccharide, or a di- or trisaccharide monomer unit of heparin,    heparan sulfate, keratin, keratan sulfate, chondroitin,    chondoitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatan    sulfate, dextran, or hyaluronic acid; and-   n is the number of lipid moiety molecules bound to a molecule of X    wherein n is a number from 1 to 1000.

In another embodiment, related low molecular weight derivatives for usein this invention are exemplified hereinabove by the general formulae(XIII) wherein:

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a mono- or disaccharide, carboxylated disaccharide, mono- or    dicarboxylic acids, a salicylate, salicylic acid, aspirin,    lactobionic acid, maltose, an amino acid, glycine, acetic acid,    butyric acid, dicarboxylic acid, glutaric acid, succinic acid, fatty    acid, dodecanoic acid, didodecanoic acid, bile acid, cholic acid,    cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, a    trisaccharide, or a di- or trisaccharide monomer unit of heparin,    heparan sulfate, keratin, keratan sulfate, chondroitin,    chondoitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatan    sulfate, dextran, or hyaluronic acid; and-   n is the number of lipid moiety molecules bound to a molecule of X    wherein n is a number from 1 to 1000.

In another embodiment, related low molecular weight derivativesaccording to the invention may be exemplified herein by any of thegeneral formulae (A), (I)-(XXI) wherein:

In one embodiment of the invention, X is covalently conjugated to alipid. In another embodiment, x is covalently conjugated to a lipid viaan amide bond. In another embodiment, x is covalently conjugated to alipid via an esteric bond. In another embodiment, the lipid isphosphatidylethanolamine. In another embodiment, the GAG may be, ininter alia, chondroitin sulfate. In another embodiment, the conjugate isbiodegradable.

In one embodiment, the invention provides glycosaminoglycans (GAG)compound covalently conjugated to a lipid to obtain a compound havingpreferred therapeutic properties. In another embodiment, the GAGcompound is covalently conjugated to a lipid via an amide bond. Inanother embodiment, the GAG compound is covalently conjugated to a lipidvia an esteric bond. In another embodiment, the lipid may be, interalia, phosphatidylethanolamine. In another embodiment, the GAG may be,inter alia, chondroitin sulfate. In another embodiment, the conjugate isbiodegradable.

Cell surface GAG play a key role in protecting cells from diversedamaging agents and processes, such as reactive oxygen species and freeradicals, endotoxins, cytokines, invasion promoting enzymes, and agentsthat induce and/or facilitate degradation of extracellular matrix andbasal membrane, cell invasiveness, white cell extravasation andinfiltration, chemotaxis, and others. In addition, cell surface GAGprotect cells from bacterial, viral and parasite infection, and theirstripping exposes the cell to interaction and subsequent internalizationof the microorganism. Enrichment of cell surface GAG would thus assistin protection of the cell from injurious processes. Thus, In oneembodiment of the invention, PLA2 inhibitos were conjugated to GAGs orGAG-mimicking molecules. In another embodiment, these Lipid-conjugates,provides wide-range protection from diverse injurious processes, and areeffective in amelioration of diseases that requires cell protection frominjurous biochemical medistors.

In another embodiment, GAG-mimicking molecule may be, inter alia, anegatively charged molecule. In another embodiment, GAG-mimickingmolecule may be, inter alia, a salicilate derivative. In anotherembodiment, GAG-mimicking molecule may be, idler alia, a dicarboxylicacid.

Preparation of Compounds

The preparation of some high molecular weight Lipid-conjugates is thesubject of U.S. Pat. No. 5,064,817, which is incorporated herein byreference. These synthetic methods are reiterated below and areconsidered to be applicable as well to the preparation of low molecular,i.e. Lipid-conjugates comprising monomers and dimers as the conjugatedmoiety, with modifications in the procedure as readily evident to oneskilled in the art.

When the starting compound chosen for the conjugated moiety has asubstituent which is or can be rendered reactive to a substituent on thestarting Lipid compound, the conjugated carrier moiety may be linkeddirectly to lipid molecule(s) to produce the a Lipid-conjugate. When itdoes not, a bifunctional linking starting material can be used to linkthe two molecules indirectly.

Lipid-conjugates are prepared by linking a polar conjugate, e.g., amonomer or polymer, directly or indirectly to a PL moiety according tothe general reaction schemes delineated in U.S. Pat. No. 5,064,817.

For example, with acylated PE used as precursor for the PE conjugate,various lengths of dicarboxylic acids can be used as spacers. Theseacids can be linked to natural, semi-synthetic or synthetic PE.

For example, PE can be linked to aminodextran indirectly as delineatedin U.S. Pat. No. 5,064,817,

Polymers with carboxylic groups, such as polyamino acids, carboxymethylcellulose or polymers to which fatty acids have been linked, can belinked directly to PE according to the scheme delineated in U.S. Pat.No. 5,064,817.

It is to be understood that these examples are given by way ofillustration only and are not to be construed as limiting the inventioneither in spirit of in scope, as many modifications both in reagents andmethods could be possible to those skilled in the art. Based on the widespectrum of pharmacological properties exhibited by Lipid-conjugates, itis likely that compounds covered by Formula I-XXI, in addition to thoseexplicitly described above, have the same valuable biological activitiesdemonstrate to be useful in the methods of treating disease describedbelow.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (A):

wherein

-   L is a lipid or a phospholipid;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein X is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between L, Z, Y and X is either an amide or an    esteric bond, including, inter alia, the steps of:    -   conjugating L to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, L is conjugated directly to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, L is conjugated directly to X,    -   thereby preparing a compound represented by the structure of the        general formula (A).

In another embodiment, the invention provides a process for thepreparation of a compound represented by the structure of the generalformula (I):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is either a physiologically acceptable monomer, dimer,        oligomer or a physiologically acceptable polymer, wherein X is a        glycosaminoglycan; and    -   n is a number from 1 to 1,000;        wherein if Y is nothing the phosphatidylethanolamine is directly        linked to X via an amide bond and if Y is a spacer, the spacer        is directly linked to X via an amide or an esteric bond and to        the phosphatidylethanolamine via an amide bond, including, inter        alia, the steps of:    -   conjugating the phosphatidylethanolamine to Y; and    -   conjugating Y to X;    -   if Y is nothing, the phosphatidylethanolamine is conjugated        directly to X,    -   thereby preparing a compound represented by the structure of the        general formula (I).

In one embodiment of the invention, the phosphatidylethanolamine is thechemical moiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In another embodiment, the invention provides a process for thepreparation of a compound represented by the structure of the generalformula (II):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein if Y is nothing the phosphatidylserine is directly linked to    X via an amide bond and if Y is a spacer, the spacer is directly    linked to X via an amide or an esteric bond and to the    phosphatidylserine via an amide bond, including, inter alia, the    steps of:    -   conjugating the phosphatidylserine to Y;    -   conjugating Y to X;    -   if Y is nothing, the phosphatidylserine is conjugated directly        to X,    -   thereby preparing a compound represented by the structure of the        general formula (II).

In one embodiment of the invention, the phosphatidylserine is thechemical moiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (III):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms:-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phosphatidyl, Z, Y and X is either an    amide or anesteric bond, including, inter alia, the steps of:    -   conjugating the phosphatidyl to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the phosphatidyl is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the phosphatidyl is conjugated directly        to X,    -   thereby preparing a compound represented by the structure of the        general formula (III).

In one embodiment of the invention, the phosphatidyl may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (IV):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond, including, inter alia, the steps of:    -   conjugating the phospholipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the phospholipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the phospholipid is conjugated directly        to X,    -   thereby preparing a compound represented by the structure of the        general formula (IV).

In one embodiment of the invention, the phospholipid may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (V):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond, including, inter, alia, the steps of:    -   conjugating the phospholipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the phospholipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the phospholipid is conjugated directly        to X,    -   thereby preparing a compound represented by the structure of the        general formula (V).

In one embodiment of the invention, the phospholipid may be the chemicalmoiety represented by the structure of

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (VI):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, inositol, choline, or glycerol;-   V is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond, including, inter alia, the steps of:    -   conjugating the phospholipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the phospholipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the phospholipid is conjugated directly        to X, thereby preparing a compound represented by the structure        of the general formula (VI).

In one embodiment of the invention, the phospholipid may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (VII):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, inositol, choline, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer; oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond, including, inter alia, the steps of:    -   conjugating the phospholipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the phospholipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the phospholipid is conjugated directly        to X, thereby preparing a compound represented by the structure        of the general formula (VII).

In one embodiment of the invention, the phospholipid may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula(VIII):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond, including, inter alia, the steps of:    -   conjugating the phospholipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the phospholipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the phospholipid is conjugated directly        to X, thereby preparing a compound represented by the structure        of the general formula (VIII).

In one embodiment of the invention, the phospholipid may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (IX):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond, including, inter alia, the steps of:    -   conjugating the phospholipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the phospholipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the phospholipid is conjugated directly        to X, thereby preparing a compound represented by the structure        of the general formula (IX).

In one embodiment of the invention, the phospholipid may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (IXa):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond, including, inter, alia, the steps of:    -   conjugating the phospholipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the phospholipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the phospholipid is conjugated directly        to X, thereby preparing a compound represented by the structure        of the general formula (IXa).

In one embodiment of the invention, the phospholipid may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (IXb):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the phospholipid, Z, Y and X is either an    amide or an esteric bond, including, inter alia, the steps of:    -   conjugating the phospholipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the phospholipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the phospholipid is conjugated directly        to X, thereby preparing a compound represented by the structure        of the general formula (IXb).

In one embodiment of the invention, the phospholipid may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (X):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the ceramide phosphoryl, Z, Y and X is    either an amide or an esteric bond, including, inter alia, the steps    of:    -   conjugating the ceramide phosphoryl to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the ceramide phosphoryl is conjugated        directly to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the ceramide phosphoryl is conjugated        directly to X, thereby preparing a compound represented by the        structure of the general formula (X).

In one embodiment of the invention, the ceramide phosphoryl may be thechemical moiety represented by the structure of:

wherein R₁ and R₂ are defined herein

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (XI):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein if Y is nothing the sphingosyl is directly linked to X via    an amide bond and if Y is a spacer, the spacer is directly linked to    X and to the sphingosyl via an amide bond and to X via an amide or    an esteric bond, including, inter alia, the steps of:    -   conjugating the sphingosyl to Y;    -   conjugating Y to X;    -   wherein if Y is nothing, the sphingosyl is conjugated directly        to X, thereby preparing a compound represented by the structure        of the general formula (XI)

In one embodiment of the invention, the sphingosyl may be the chemicalmoiety represented by the structure of:

wherein R₁ is defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (XII):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   L is ceramide;-   Z is either nothing, ethanolamine, serine, inositol, choline, or    glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the ceramide, Z, Y and X is either an amide    or an esteric bond, including, inter alia, the steps of:    -   conjugating the ceramide to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the ceramide is conjugated directly to        Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the ceramide is conjugated directly to        X, thereby preparing a compound represented by the structure of        the general formula (XII).

In one embodiment of the invention, the ceramide may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula(XIII):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the diglyceryl, Z, Y and X is either an    amide or an esteric bond, including, inter alia, the steps of:    -   conjugating the diglyceryl to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the diglyceryl is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the diglyceryl is conjugated directly to        X, thereby preparing a compound represented by the structure of        the general formula (XIII).

In one embodiment of the invention, the diglyceryl may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (XIV):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer; oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the glycerolipid, Z, Y and X is either an    amide or an esteric bond, including, inter alia, the steps of:    -   conjugating the glycerolipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the glycerolipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the glycerolipid is conjugated directly        to X, thereby preparing a compound represented by the structure        of the general formula (XIV).

In one embodiment of the invention, the glycerolipid may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (XV):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the glycerolipid, Z, Y and X is either an    amide or an esteric bond, including, inter alia, the steps of:    -   conjugating the glycerolipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the glycerolipid is conjugated directly        to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the glycerolipid is conjugated directly        to X, thereby preparing a compound represented by the structure        of the general formula (XV).

In one embodiment of the inventions the glycerolipid may be the chemicalmoiety represented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (XVI):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond, including, inter alia, the steps of:    -   conjugating the lipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the lipid is conjugated directly to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the lipid is conjugated directly to X,        thereby preparing a compound represented by the structure of the        general formula (XVI).

In one embodiment of the invention, the lipid may be the chemical moietyrepresented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula(XVII):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond, including, inter alia, the steps of:    -   conjugating the lipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the lipid is conjugated directly to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the lipid is conjugated directly to X,        thereby preparing a compound represented by the structure of the        general formula (XVII).

In one embodiment of the invention, the lipid may be the chemical moietyrepresented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula(XVIII):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond, including, in inter alia, the steps of:    -   conjugating the lipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the lipid is conjugated directly to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the lipid is conjugated directly to X,        thereby preparing a compound represented by the structure of the        general formula (XVIII)

In one embodiment of the invention, the lipid may be the chemical moietyrepresented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (XIX):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond, including, inter alia, the steps of:    -   conjugating the lipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the lipid is conjugated directly to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the lipid is conjugated directly to X,        thereby preparing a compound represented by the structure of the        general formula (XIX)

In one embodiment of the invention, the lipid may be the chemical moietyrepresented by the structure of:

wherein R₁ and R₂ are defined herein

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (XX):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond, including, inlet alia, the steps of:    -   conjugating the lipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the lipid is conjugated directly to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the lipid is conjugated directly to X,        thereby preparing a compound represented by the structure of the        general formula (XX).

In one embodiment of the invention, the lipid may be the chemical moietyrepresented by the structure of:

wherein R₁ and R₂ are defined herein.

In one embodiment, the invention provides a process for the preparationof a compound represented by the structure of the general formula (XXI):

wherein

-   R₁ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   R₂ is either hydrogen or a linear, saturated, mono-unsaturated, or    poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon    atoms;-   Z is either nothing, choline, phosphate, inositol, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer, wherein x is a glycosaminoglycan; and-   n is a number from 1 to 1000;    wherein any bond between the lipid, Z, Y and X is either an amide or    an esteric bond, including, inter alia, the steps of:    -   conjugating the lipid to Z;    -   conjugating Z to Y;    -   conjugating Y to X;    -   wherein if Z is nothing, the lipid is conjugated directly to Y,    -   if Y is nothing, Z is conjugated directly to X, and    -   if Y and Z are nothing, the lipid is conjugated directly to X,        thereby preparing a compound represented by the structure of the        general formula (XXI).

In one embodiment of the invention, the lipid may be the chemical moietyrepresented by the structure of:

wherein R₁ and R₂ are defined herein.

In another embodiment, the conjugating according to the invention, maybe performed by eliminating a water molecule, thereby forming amide oresteric bonds. In another embodiment, the conjugating may be performedin the presence of a detergent. In another embodiment, the conjugatingmay be induced by ultrasonic radiation.

In another embodiment, any conjugation process according to theinvention may be performed by eliminating a water molecule, therebyforming amide or esteric bonds. In another embodiment, any conjugationprocess according to the invention may be performed in the presence of adetergent. In another embodiment, any conjugation process according tothe invention may be induced by ultrasonic radiation.

In another embodiment, any compound according to the invention may beprepared by a conjugation process performed by eliminating a watermolecule, thereby forming amide or esteric bonds. In another embodiment,any compound according to the invention may be prepared by a conjugationprocess in the presence of a detergent. In another embodiment, anycompound according to the invention may be prepared by a conjugationprocess induced by ultrasonic radiation.

In one embodiment of the invention, the conjugation of thephosphatidylethanolamine and chondroitin sulfate is performed in thepresence of a detergent. In another embodiment a detergent may be, interalia, DDAB Of course any other appropriate detergent may be used.

In one embodiment of the invention, the conjugation of thephosphatidylethanolamine and hyaluronic acid is iduced by sonication.

Methods of Treating Disease Based on PL Conjugates

In one embodiment of the invention, the Lipid-conjugates describedherein can be used to treat disease, through exerting at least one oftheir many pharmacological activities, among which are amelioration, orprevention, of tissue injury arising in the course of pathologicaldisease states by stabilizing cell membranes; limiting oxidative damageto cell and blood components; limiting cell proliferation, cellextravasation and (tumor) cell migratory behavior, suppressing immuneresponses; or attenuating physiological reactions to stress, asexpressed in elevated chemokine levels. The medicinal properties ofthese compounds are readily exemplified in using animal models of theparticular disease in which it is desired to use the drug. The patientsto whom the lipid or PL conjugates should be administered are those thatare experiencing symptoms of disease or who are at risk of contractingthe disease or experiencing a recurrent episode or exacerbation of thedisease. The efficacy of these compounds in cellular and animal modelsof disease are described below in The Examples.

The combination of lipids, such as, but not limited tophosphatidylethanolamine and phosphatidylserine, with additional monomeror polymer moieties, is thus a practical route to die production of newdrugs for medical purposes, provided that the resultant chemicalcomposition displays the desired range of pharmacological properties. Inthe cases described herein, the diversity of biological activities andthe effectiveness in disease exhibited by the compounds far exceed theproperties anticipated by use of the starting materials themselves, whenadministered alone or in combination. However, it is likely that the PLconjugate compounds, alone or in combination, will prove to be valuabledrugs when adapted to methods of disease treatment other to thoseconditions specifically described herein.

In one embodiment, the invention provides a method of treating a subjectafflicted with a disease related to asthma, rhinitis, allergic rhinitis,chronic obstructive pulmonary disease, obstructive respiratory disease,chlamydia infection, a disorder of smooth muscle cell proliferation,metastatic cancer, colitis, Crohn's disease, or another form ofintestinal mucosal injury, cardiovascular disease, atherosclerosis,central nervous system tissue insult, multiple sclerosis, contactdermatitis, psoriasis, cellular proliferative disorder, sepsis, acuterespiratory distress syndrome, autoimmune disease, hemolysis, HIVinfection, or conjunctivitis.

In one embodiment, the invention provides a method of treating a subjectsuffering from asthma, including, idler alia, the step of administeringto a subject an effective amount of a lipid or phospholipid moietybonded to a physiologically acceptable monomer, dimer, oligomer, orpolymer.

In one embodiment, the invention provides a method of preventing asthmain a subject, including, inter alia, the step of administering to asubject an effective amount of a lipid OT phospholipid moiety bonded toa physiologically acceptable monomer, dimer, oligomer, or polymer.

In one embodiment, the invention provides a method of treating a subjectsuffering from allergic rhinitis, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer.

In one embodiment, the invention provides a method of preventingallergic rhinitis in a subject, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer.

In one embodiment, the invention provides a method of treating a subjectsuffering from chronic obstructive pulmonary disease, including, interalia, the step of administering to a subject an effective amount of alipid or phospholipid moiety bonded to a physiologically acceptablemonomer, dimer, oligomer, or polymer.

In one embodiment, the invention provides a method of preventing chronicobstructive pulmonary disease in a subject, including, inter alia, thestep of administering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer.

In one embodiment, the invention provides a method of treating a subjectsuffering from obstructive respiratory disease, including, inter alia,the step of administering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer.

In one embodiment, the invention provides a method of preventingobstructive respiratory disease in a subject, including, inter alia, thestep of administering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer.

In one embodiment, the invention provides a method of treating a subjectrequiring anti-oxidant therapy, including, infer alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject requiring ananti-oxidant therapy.

In one embodiment, the invention provides a method treating a subjectrequiring anti-TNF therapy, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject requiring ananti-TNF therapy.

In one embodiment, the invention provides a method of treating a subjectsuffering from a disorder of smooth muscle cell proliferation,including, inter alia, the step of administering to a subject aneffective amount of a lipid or phospholipid moiety bonded to aphysiologically acceptable monomer, dimer, oligomer, or polymer, therebytreating the subject suffering from a disorder related to smooth musclecell proliferation

In one embodiment, the invention provides a method of treating a subjectundergoing vascular catheterization, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject undergoingvascular catheterization.

In one embodiment, the invention provides a method of treating a subjectsuffering from metastatic cancer, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject suffering frommetastatic cancer.

In one embodiment, the invention provides a method of treating a subjectsuffering from colitis, Crohn's disease, or another form of intestinalmucosal injury, including, in inter alia, the step of administering to asubject an effective amount of a lipid or phospholipid moiety bonded toa physiologically acceptable monomer, dimer, oligomer, or polymer,thereby treating the subject suffering from intestinal mucosal injury,including colitis or Crohn's disease.

In one embodiment, the invention provides a method of treating a subjectsuffering from cardiovascular disease, including, inter alia, the stepof administering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject suffering froma cardiovascular disease.

The present invention provides a method of treating a subject sufferingfrom atherosclerosis, including, inter alia, the step of administeringto a subject an effective amount of a lipid or phospholipid moietybonded to a physiologically acceptable monomer, dimer, oligomer, orpolymer, thereby treating the subject suffering from atherosclerosis.

In one embodiment, the invention provides a method of treating a subjectsuffering from central nervous system tissue insult, including, interalia, the step of administering to a subject an effective amount of alipid or phospholipid moiety bonded to a physiologically acceptablemonomer, dimer; oligomer, or polymer, thereby treating the subjectsuffering from a central nervous system insult.

In one embodiment, the invention provides a method of treating a subjectsuffering from multiple sclerosis, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject suffering frommultiple sclerosis.

In one embodiment, the invention provides a method of treating a subjectsuffering from contact dermatitis, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject suffering fromcontact dermatitis.

In one embodiment, the invention provides a of treating a subjectsuffering from psoriasis, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject suffering frompsoriasis.

In one embodiment, the invention provides a method of treating a subjectsuffering from a cellular proliferative disorder, including, inter alia,the step of administering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject suffering froma cellular proliferative disorder.

In one embodiment, the invention provides a method of treating a subjectsuffering from sepsis, including, inter alia, the step of administeringto a subject an effective amount of a lipid or phospholipid moietybonded to a physiologically acceptable monomer, dimer, oligomer, orpolymer, thereby treating the subject suffering from sepsis.

In one embodiment, the invention provides a method of treating a subjectsuffering from ARDS, comprising the steps of administering to a subjectan effective amount of a lipid or phospholipid moiety bonded to aphysiologically acceptable monomer, dimer, oligomer, or polymer, therebytreating the subject suffering from ARDS.

In one embodiment, the invention provides a method of treating a subjectsuffering from autoimmune disease, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject suffering froman autoimmune disease.

In one embodiment, the invention provides a method of treating a subjectsuffering from hemolysis, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject suffering fromhemolysis.

In one embodiment, the invention provides a method of treating a subjectundergoing tissue transplantation or allograft rejection, including,inter alia, the step of administering to a subject an effective amountof a lipid or phospholipid moiety bonded to a physiologically acceptablemonomer, dimer, oligomer, or polymer, thereby treating the subjectundergoing tissue transplantation or allograft rejection.

In one embodiment, the invention provides a method of treating a subjectafflicted with HIV infection, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject afflicted withHIV infection.

In one embodiment, the invention provides a method of treating a subjectafflicted with conjunctivitis, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject afflicted withconjunctivitis.

In one embodiment, the invention provides a method for extracorporealtissue preservation, including, inter alia, the step of adding to atissue preparation or organ an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby extending the viability of thetissue preparation or organ within a donor subject.

In one embodiment, the invention provides a method of treating a subjectafflicted with Chlamydia infection, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject afflictedsuffering from Chlamydia infection.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from asthma.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for preventing asthma in a subject.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from allergic rhinitis.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for preventing allergic rhinitis in a subject.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from chronic obstructivepulmonary disease.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for preventing chronic obstructive pulmonary disease in asubject.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from obstructiverespiratory disease.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for preventing obstructive respiratory disease in a subject.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject requiring an anti-oxidant therapy.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject requiring an anti-TNF therapy.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer- or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from a disorder related tosmooth muscle cell proliferation.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject undergoing vascular catheterization.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering form metastatic cancer.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from intestinal mucosalinjury, including inter alia, colitis or Crohn's disease.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer in the preparation of a pharmaceuticalcomposition for treating a subject suffering from a cardiovasculardisease.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from atherosclerosis.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for, treating a subject suffering from central nervoussystem insult.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from multiple sclerosis.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from contact dermatitis.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from psoriasis.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from a cellularproliferative disorder.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from sepsis.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer; oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from ARDS.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from an autoimmune disease.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject suffering from hemolysis.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject undergoing tissue transplantation orallograft rejection.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject afflicted with HIV infection.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject afflicted with conjunctivitis.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for extending the viability of the tissue preparation ororgan within a donor subject.

In one embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject afflicted with Chlamydia infection.

In one embodiment of the invention, the treatment requires controllingthe expression production and activity of phospholipase enzymes. Inanother embodiment, the treatment requires controlling the productionand/or action of lipid mediators. In another embodiment, the treatmentrequires amelioration of damage to glycosaminoglycans (GAG) andproteoglycans. In another embodiment, the treatment requires controllingthe production and action of oxidants, oxygen radicals and nitric oxide.In another embodiment, the treatment requires anti-oxidant therapy. Inanother embodiment, the treatment requires anti-endotoxin therapy. Inanother embodiment, the treatment requires controlling the expression,production or action of cytokines, chemokines, adhesion molecules orinterleukines. In another embodiment, the treatment requires protectionof lipoproteins from damaging agents. In another embodiment, thetreatment requires controlling the proliferation of cells. In anotherembodiment, the treatment requires controlling of angiogenesis and organvascularization. In another embodiment, the treatment requiresinhibition of invasion-promoting enzymes. In another embodiment, thetreatment requires controlling of cell invasion. In another embodiment,the invading cells are white blood cells. In another embodiment, theinvading cells are cancer cells. In another embodiment, the treatmentrequires controlling of white cell activation, adhesion orextravasation. In another embodiment, the treatment requiresamelioration of ischemia or reperfusion injury. In another embodiment,the treatment requires inhibition of lymphocyte activation. In anotherembodiment, the treatment requires protection of blood brain barrier. Inanother embodiment, the treatment requires control of neurotransmitterproduction and action. In another embodiment, the treatment requirescontrolling of blood vessel and airway contraction. In anotherembodiment, the treatment requires extracorporeal tissue preservation.

In one embodiment of the invention, the lipid mediator is aglycerolipid. In another embodiment, the lipid mediator is aphospholipid. In another embodiment, the lipid mediator is sphingolipid.In another embodiment, the lipid mediator is a sphingosine. In anotherembodiment, the lipid mediator is ceramide. In another embodiment, thelipid mediator is a fatty acid. In another embodiment, the fatty acid isarachidonic acid. In another embodiment, the lipid mediator is anarachidonic acid-derived eicosanoid. In another embodiment, the lipid ismediator is a platelet activating factor (PAF). In another embodiment,the lipid mediator is a lysophospholipid.

In one embodiment of the invention, the damaging agent is aphospholipase. In another embodiment, the damaging agent is a reactiveoxygen species (ROS). In another embodiment, the damaging agent is afree radical. In another embodiment, the damaging agent is alysophospholipid. In another embodiment, the damaging agent is a fattyacid or a derivative thereof. In another embodiment, the damaging agentis hydrogen peroxide. In another embodiment, the damaging agent is aphospholipid. In another embodiment, the damaging agent is an oxidant.In another embodiment, the damaging agent is a cationic protein. Inanother embodiment, the damaging agent is a streptolysin. In anotherembodiment, the damaging agent is a protease. In another embodiment, thedamaging agent is a hemolysin. In another embodiment, the damaging agentis a sialidase.

In one embodiment of the invention, the invasion-promoting enzyme iscollagenase. In another embodiment, the invasion-promoting enzyme ismatrix-metaloproteinase (MMP). In another embodiment, theinvasion-promoting enzyme is heparinase. In another embodiment, theinvasion-promoting enzyme is heparanase. In another embodiment, theinvasion-promoting enzyme is hyaluronidase. In another embodiment, theinvasion-promoting enzyme is gelatinase. In another embodiment, theinvasion-promoting enzyme is chondroitinase. In another embodiment, theinvasion-promoting enzyme is dermatanase. In another embodiment, theinvasion-promoting enzyme is keratanase. In another embodiment, theinvasion-promoting enzyme is protease. In another embodiment, theinvasion-promoting enzyme is lyase. In another embodiment, theinvasion-promoting enzyme is hydrolase. In another embodiment, theinvasion-promoting enzyme is a glycosaminoglycan degrading enzyme. Inanother embodiment, the invasion-promoting enzyme is a proteoglycandegrading enzyme.

In one embodiment of the invention, the physiologically acceptablemonomer is salicylate. In another embodiment, the physiologicallyacceptable monomer is salicylic acid. In another embodiment, thephysiologically acceptable monomer is aspirin. In another embodiment,the physiologically acceptable monomer is a monosaccharide. In anotherembodiment, the physiologically acceptable monomer is lactobionic acid.In another embodiment, the physiologically acceptable monomer isglucoronic acid. In another embodiment, the physiologically acceptablemonomer is maltose. In another embodiment, the physiologicallyacceptable monomer is an amino acid. In another embodiment, thephysiologically acceptable monomer is glycine. In another embodiment,the physiologically acceptable monomer is a carboxylic acid. In anotherembodiment, the physiologically acceptable monomer is an acetic acid. Inanother embodiment, the physiologically acceptable monomer is a butyricacid. In another embodiment, the physiologically acceptable monomer is adicarboxylic acid. In another embodiment, the physiologically acceptablemonomer is a glutaric acid. In another embodiment, the physiologicallyacceptable monomer is succinic acid. In another embodiment, thephysiologically acceptable monomer is a fatty acid. In anotherembodiment, the physiologically acceptable monomer is dodecanoic acid.In another embodiment, the physiologically acceptable monomer isdidodecanoic acid. In another embodiment, the physiologically acceptablemonomer is bile acid. In another embodiment, the physiologicallyacceptable monomer is cholic acid. In another embodiment, thephysiologically acceptable monomer is cholesterylhemmisuccinate.

In one embodiment of the invention, the physiologically acceptable dimeror oligomer is physiologically acceptable dimer or oligomer is adipeptide. In another embodiment, the physiologically acceptable dimeror oligomer is a disaccharide. In another embodiment, thephysiologically acceptable dimer or oligomer is a trisaccharide. Inanother embodiment, the physiologically acceptable dimer or oligomer isan oligosaccharide. In another embodiment, the physiologicallyacceptable dimer or oligomer is an oligopeptide. In another embodiment,the physiologically acceptable dimer or oligomer is a di- ortrisaccharide monomer unit of glycosaminoglcans. In another embodiment,the physiologically acceptable dimer or oligomer is hyaluronic acid. Inanother embodiment, the physiologically acceptable dimer or oligomer isheparin. In another embodiment, the physiologically acceptable dimer oroligomer is heparan sulfate. In another embodiment, the physiologicallyacceptable dimer or oligomer is keratin. In another embodiment, thephysiologically acceptable dimer or oligomer is keratan sulfate. Inanother embodiment, the physiologically acceptable dimer or oligomer ischondroitin. In another embodiment, the chondroitin is chondoitinsulfate. In another embodiment, the chondroitin is chondoitin-4-sulfate.In another embodiment, the chondroitin is chondoitin-6-sulfate. Inanother embodiment, the physiologically acceptable dimer or oligomer isdermatin. In another embodiment, the physiologically acceptable dimer oroligomer is dermatan sulfate. In another embodiment, the physiologicallyacceptable dimer or oligomer is dextran. In another embodiment, thephysiologically acceptable dimer or oligomer is polygeline(‘Haemaccel’). In another embodiment, the physiologically acceptabledimer or oligomer is alginate. In another embodiment, thephysiologically acceptable dimer or oligomer is hydroxyethyl starch(Hetastarch). In another embodiment, the physiologically acceptabledimer or oligomer is ethylene glycol. In another embodiment, thephysiologically acceptable dimer or oligomer is carboxylated ethyleneglycol.

In one embodiment of the invention, the physiologically acceptablepolymer is a glycosaminoglycan. In another embodiment, thephysiologically acceptable polymer is hyaluronic acid. In anotherembodiment, the physiologically acceptable polymer is heparin. Inanother embodiment, the physiologically acceptable polymer is heparansulfate. In another embodiment, the physiologically acceptable polymeris chondroitin. In another embodiment, the chondroitin ischondoitin-4-sulfate. In another embodiment, the chondroitin ischondoitin-6-sulfate. In another embodiment, the physiologicallyacceptable polymer is keratin. In another embodiment, thephysiologically acceptable polymer is keratan sulfate. In anotherembodiment, the physiologically acceptable polymer is dermatin. Inanother embodiment, the physiologically acceptable polymer is dermatansulfate. In another embodiment, the physiologically acceptable polymeris carboxymethylcellulose. In another embodiment, the physiologicallyacceptable polymer is dextran. In another embodiment, thephysiologically acceptable polymer is polygeline (‘Haemaccel’). Inanother embodiment, the physiologically acceptable polymer is alginate.In another embodiment, the physiologically acceptable polymer ishydroxyethyl starch (‘Hetastarch’). In another embodiment, thephysiologically acceptable polymer is polyethylene glycol. In anotherembodiment, the physiologically acceptable polymer is polycarboxylatedpolyethylene glycol.

In one embodiment of the invention, the lipid or phospholipid moiety isphosphatidic acid. In another embodiment, lipid or phospholipid moietyis an acyl glycerol. In another embodiment, lipid or phospholipid moietyis monoacylglycerol. In another embodiment, lipid or phospholipid moietyis diacylglycerol. In another embodiment, lipid or phospholipid moietyis triacylglycerol. In another embodiment, lipid or phospholipid moietyis sphingosine. In another embodiment, lipid or phospholipid moiety issphingomyelin. In another embodiment, lipid or phospholipid moiety isceramide. In another embodiment, lipid or phospholipid moiety isphosphatidylethanolamine. In another embodiment, lipid or phospholipidmoiety is phosphatidylserine. In another embodiment, lipid orphospholipid moiety is phosphatidylcholine. In another embodiment, lipidor phospholipid moiety is phosphatidylinositol. In another embodiment,lipid or phospholipid moiety is phosphatidylglycerol. In anotherembodiment, lipid or phospholipid moiety is an ether or alkylphospholipid derivative thereof

In one embodiment, the invention provides a method of treating a subjectafflicted with a disease, wherein the treatment of the disease requirescontrolling phospholipase A2 activities; controlling the productionand/or action of lipid mediators, such as eicosanoids, plateletactivating factor (PAF) and lyso-phospholipids; amelioration of damageto cell surface glycosaminoglycans (GAG) and proteoglycans; controllingthe production of oxygen radicals and nitric oxide; protection of cells,tissues, and plasma lipoproteins from damaging agents, such as reactiveoxygen species (ROS) and phospholipases; anti-oxidant therapy;anti-endotoxin therapy; controlling of cytokine, chemokine andinterleukine production; controlling the proliferation of cells,including smooth muscle cells, endothelial cells and skin fibroblasts;controlling of angiogenesis and organ vascularization; inhibition ofinvasion-promoting enzymes, such as collagenase, heparinase, heparanaseand hyaluronidase; controlling of cell invasion; controlling of whitecell activation, adhesion and extravasation; amelioration ofischemia/reperfusion injury, inhibition of lymphocyte activation;controlling of blood vessel and airway contraction; protection of bloodbrain barrier; controlling of neurotransmitter (e.g., dopamine)production and action (e.g., acethylcholine); extracorporeal tissuepreservation or any combination thereof

In one embodiment of the invention, the term “controlling” refers toinhibiting the production and action of the above mentioned factors inorder to maintain their activity at the normal basal level and suppresstheir activation in pathological conditions.

In one embodiment of the invention, the physiologically acceptablemonomer is either a salicylate, salicylic acid, aspirin, amonosaccharide, lactobionic acid, maltose, an amino acid, glycine,carboxylic acid, acetic acid, butyric acid, dicarboxylic acid, glutaricacid, succinic acid, fatty acid, dodecanoic acid, didodecanoic acid,bile acid, cholic acid, cholesterylhemmisuccinate; or wherein thephysiologically acceptable dimer or oligomer is a dipeptide, adisaccharide, a trisaccharide, an oligopeptide, or a di- ortrisaccharide monomer unit of heparin, heparan sulfate, keratin, keratansulfate, chondroitin, chondoitin-6-sulfate, chondroitin-4-sulfate,dermatin, dermatan sulfate, dextran, or hyaluronic acid; or wherein thephysiologically acceptable polymer is a glycosaminoglycan, polygelin(‘haemaccel’), alginate, hydroxyethyl starch (hetastarch), polyethyleneglycol, polycarboxylated polyethylene glycol, chondroitin-6-sulfate,chondroitin-4-sulfate, keratin, keratin sulfate, heparan sulfate,dermatin, dermatan sulfate, carboxymethylcellulose, heparin, dextran, orhyaluronic acid.

In one embodiment of the invention, the lipid moiety is eitherphosphatidic acid, an acyl glycerol, monoacylglycerol, diacylglycerol,triacylglycerol, sphingosine, sphingomyelin, chondroitin-4-sulphate,chondroitin-6-sulphate, ceramide, phosphatidylethanolamine,phosphatidylserine, phosphatidylcholine, phosphatidylinositol, orphosphatidylglycerol, or an ether or alkyl phospholipid derivativethereof, and the physiologically acceptable monomer or polymer moiety iseither aspirin, lactobionic acid, maltose, glutaric acid, polyethyleneglycol, carboxymethylcellulose, heparin, dextran, hemacell, hetastarch,or hyaluronic acid.

In one embodiment, the present invention provides for use of a lipidmoiety bonded to a physiologically acceptable monomer, dimer, oligomer,or polymer, in the preparation of a pharmaceutical composition fortreating a subject afflicted with asthma, allergic rhinitis, chronicobstructive pulmonary disease, obstructive respiratory disease, colitis,Crohn's disease, central nervous system insult, multiple sclerosis,contact dermatitis, psoriasis, cardiovascular disease, includingprophylaxis for invasive procedures, invasive cellular proliferativedisorders, anti-oxidant therapy, hemolytic syndromes, sepsis, acuterespiratory distress syndrome, tissue transplant rejection syndromes,autoimmune disease, viral infection, and hypersensitivityconjunctivitis.

In one embodiment, the present invention provides use of apharmaceutical composition according to the present invention fortreating a subject afflicted with asthma, allergic rhinitis, chronicobstructive pulmonary disease, obstructive respiratory disease, colitis,Crohn's disease, central nervous system insult, multiple sclerosis,contact dermatitis, psoriasis, cardiovascular disease, includingprophylaxis for invasive procedures, invasive cellular proliferativedisorders, anti-oxidant therapy, hemolytic syndromes, sepsis, acuterespiratory distress syndrome, tissue transplant rejection syndromes,autoimmune disease, viral infection, or hypersensitivity conjunctivitis,wherein the composition is prepared for administration by topical, oral,nasal, aerosol, intravenous, intraocular, intra-arterial, subcutaneous,or suppository routes.

In one embodiment, the invention provides a method of treating a subjectsuffering from an intestinal disease, including, inter alia, the step ofadministering to a subject an effective amount of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, thereby treating the subject suffering froman intestinal disease

In another embodiment, the invention provides a use of a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer, in the preparation of a pharmaceuticalcomposition for treating a subject afflicted with an intestinal disease.

In one embodiment, the invention provides a method of treating a subjectsuffering from a disease involving the production and/or action of lipidmediators and/or impairment of glycosaminoglycan (GAG) functioning.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from an intestinal disease,including, inter alia, a lipid or phospholipid moiety bonded to aphysiologically acceptable monomer, dimer, oligomer, or polymer; and apharmaceutically acceptable carrier or excipient.

In one embodiment, the intestinal disease may be, infer alia, a diseaseinvolving the production and/or action of lipid mediators and/orimpairment of glycosaminoglycan (GAG) functioning.

In one embodiment of the invention, the intestinal disease may be, interalia, Crohn's disease, ulcerative colitis, immuno-inflammatoryintestinal injury, drug-induced enteropathy, ischemia-induced intestinalinjury or any combination thereof.

In one embodiment of the invention, the physiologically acceptablemonomer may be, inter alia, a salicylate, salicylic acid, aspirin, amonosaccharide, lactobionic acid, glucoronic acid, maltose, amino acid,glycine, carboxylic acid, acetic acid, butyric acid, dicarboxylic acid,glutaric acid, succinic acid, fatty acid, dodecanoic acid, didodecanoicacid, bile acid, cholic acid, cholesterylhemmisuccinate, or wherein thephysiologically acceptable dimer or oligomer may be, inter alia, adipeptide, a disaccharide, a trisaccharide, an oligosaccharide, anoligopeptide, or a di- or trisaccharide monomer unit ofglycosaminoglcans, hyaluronic acid, heparin, heparan sulfate, keratin,keratan sulfate, chondroitin, chondroitin sulfate,chondroitin-4-sulfate, chondoitin-6-sulfate, dermatin, dermatan sulfate,dextran, polygeline, alginate, hydroxyethyl starch, ethylene glycol, orcarboxylated ethylene glycol, or wherein the physiologically acceptablepolymer may be, inter alia, a glycosaminoglycan, hyaluronic acid,heparin, heparan sulfate, chondroitin, chondroitin sulfate, keratin,keratan sulfate, dermatin, dermatan sulfate, carboxymethylcellulose,dextran, polygeline, alginate, hydroxyethyl starch, polyethylene glycolor polycarboxylated polyethylene glycol.

In another embodiment, the physiologically acceptable polymer may be,inter alia, hyaluronic acid.

In another embodiment, the physiologically acceptable polymer may be,inter alia, chondroitin sulfate.

In one embodiment of the invention, the lipid or phospholipid moiety maybe, inter alia, phosphatidic acid, an acyl glycerol, monoacylglycerol,diacylglycerol, triacylglycerol, sphingosine, sphingomyelin, ceramide,phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine,phosphatidylinositol, phosphatidylglycerol, or an ether or alkylphospholipid derivative thereof.

In another embodiment, the phospholipid moiety may be, inter alia,phosphatidylethanolamine.

Dosages and Routes of Administration

The methods of this invention can be adapted to use of the therapeuticcompositions comprising Lipid-conjugates in admixture with conventionalexcipients, i.e. pharmaceutically acceptable organic or inorganiccarrier substances suitable for parenteral, enteral (e.g., oral) ortopical application which do not deleteriously react with the activecompounds. Suitable pharmaceutically acceptable carriers include but arenot limited to water, salt solutions, alcohols, gum arabic, vegetableoils, benzyl alcohols, polyethylene glycols, gelatine, carbohydratessuch as lactose, amylose or starch, magnesium stearate, talc, silicicacid, viscous paraffin, white paraffin, glycerol, alginates, hyaluronicacid, collagen, perfume oil, fatty acid monoglycerides and diglycerides,pentaerythritol fatty acid esters, hydroxy methylcellulose, polyvinylpyrrolidone, etc. The pharmaceutical preparations can be sterilized andif desired mixed with auxiliary agents, e.g., lubricants, preservatives,stabilizers, wetting agents, emulsifiers, salts for influencing osmoticpressure, buffers, coloring, flavoring and/or aromatic substances andthe like which do not deleteriously react with the active compounds.They can also be combined where desired with other active agents, e.g.,vitamins.

In one embodiment, the invention provides a pharmaceutical compositionfor treating a subject suffering from sepsis, including a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer; and a pharmaceutically acceptable carrieror excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from asthma, including alipid or phospholipid moiety bonded to a physiologically acceptablemonomer, dimer, oligomer, or polymer; and a pharmaceutically acceptablecarrier or excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for preventing asthma in a subject, including a lipid orphospholipid moiety bonded to a physiologically acceptable monomer,dimer, oligomer, or polymer; and a pharmaceutically acceptable carrieror excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from allergic rhinitis,including a lipid or phospholipid moiety bonded to a physiologicallyacceptable monomer, dimer, oligomer, or polymer; and a pharmaceuticallyacceptable carrier or excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for preventing allergic rhinitis in a subject, including alipid or phospholipid moiety bonded to a physiologically acceptablemonomer, dimer, oligomer, or polymer; and a pharmaceutically acceptablecarrier or excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from chronic obstructivepulmonary disease, including a lipid or phospholipid moiety bonded to aphysiologically acceptable monomer, dimer, oligomer, or polymer; and apharmaceutically acceptable carrier or excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for preventing chronic obstructive pulmonary disease in asubject, including a lipid or phospholipid moiety bonded to aphysiologically acceptable monomer, dimer, oligomer, or polymer; and apharmaceutically acceptable carrier or excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from an obstructiverespiratory disease, including a lipid or phospholipid moiety bonded toa physiologically acceptable monomer, dimer, oligomer, or polymer; and apharmaceutically acceptable carrier or excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for preventing obstructive respiratory disease in a subject,including a lipid or phospholipid moiety bonded to a physiologicallyacceptable monomer, dimer, oligomer, or polymer; and a pharmaceuticallyacceptable carrier or excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from asthma, including anyone of the compounds according to the invention or any combinationthereof; and a pharmaceutically acceptable carrier or excipient. Inanother embodiment, the invention provides a pharmaceutical compositionfor preventing asthma in a subject, including any one of the compoundsaccording to the invention or any combination thereof; and apharmaceutically acceptable carrier or excipient. In another embodiment,the invention provides a pharmaceutical composition for treating asubject suffering from allergic rhinitis, including any one of thecompounds according to the invention or any combination thereof; and apharmaceutically acceptable carrier or excipient. In another embodiment,the invention provides a pharmaceutical composition for preventingallergic rhinitis in a subject, including any one of the compoundsaccording to the invention or any combination thereof; and apharmaceutically acceptable carrier or excipient. In another embodiment,the invention provides a pharmaceutical composition for treating asubject suffering from chronic obstructive pulmonary disease, includingany one of the compounds according to the invention or any combinationthereof, and a pharmaceutically acceptable carrier or excipient. Inanother embodiment, the invention provides a pharmaceutical compositionfor preventing chronic obstructive pulmonary disease in a subject,including any one of the compounds according to the invention or anycombination thereof; and a pharmaceutically acceptable carrier orexcipient. In another embodiment, the invention provides apharmaceutical composition for treating a subject suffering fromobstructive respiratory disease, including any one of the compoundsaccording to the invention or any combination thereof; and apharmaceutically acceptable carrier or excipient. In another embodiment,the invention provides a pharmaceutical composition for preventingobstructive respiratory disease in a subject, including any one of thecompounds according to the invention or any combination thereof; and apharmaceutically acceptable carrier or excipient.

In another embodiment, the compounds according to the invention include,inter alia, the compounds represented by the structures of the generalformulae: (A), (I), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX),(IXa), (IXb), (X), (XI), (XII), (XIII), (XIV), (XV), (XVI), (XVII),(XVIII), (XIX), (XX), (XXI), (XXII) or any combination thereof.

While the examples provided herein describe use of the PL conjugates insubcutaneous, intraperitoneal or topical administration the successdescribed affords good evidence to suppose that other routes ofadministration, or combinations with other pharmaceutical preparations,would be at least as successful. The route of administration (e.g.,topical, par-enteral, enteral, intravenous, vaginal, inhalation, nasalaspiration (spray), supository or oral) and the dosage regimen will bedetermined by skilled clinicians, based on factors such as exact natureof the condition being treated, the severity of the condition, the ageand general physical condition of the patient, and so on.

In general, the doses utilized for the above described purposes willvary, but will be in an effective amount to exert the desiredanti-disease effect. As used herein, the term “pharmaceuticallyeffective amount” refers to an amount of a compound of formulae A andI-XXI which will produce the desired alleviation in symptoms or signs ofdisease in a patient. The doses utilized for any of the above-describedpurposes will generally be from 1 to about 1000 milligrams per kilogramof body weight (mg/kg), administered one to four times per day, or bycontinuous IV infusion. When the compositions are dosed topically, theywill generally be in a concentration range of from 0.1 to about 10% w/N,administered 1-4 times per day.

As used herein, the term “pharmaceutically acceptable carrier” refers toany formulation which is safe, and provides the appropriate delivery forthe desired route of administration of an effective amount of at leastone compound of the present invention. As such, all of theabove-described formulations of the present invention are herebyreferred to as “pharmaceutically acceptable carriers.” This term refersto as well the use of buffered formulations wherein the pH is maintainedat a particular desired value, ranging from pH 4.0 to pH 9.0, inaccordance with is the stability of the compounds and route ofadministration.

For parenteral application, particularly suitable are injectable,sterile solutions, preferably oily or aqueous solutions, as well assuspensions, emulsions, or implants, including suppositories. Ampoulesare convenient unit dosages.

For application by inhalation, particularly for treatment of airwayobstruction or congestion, solutions or suspensions of the compoundsmixed and aerosolized or nebulized in the presence of the appropriatecarrier suitable.

For topical application, particularly for the treatment of skin diseasessuch as contact dermatitis or psoriasis, admixture of the compounds withconventional creams or delayed release patches is acceptable.

For enteral application, particularly suitable are tablets, dragees,liquids, drops, suppositories, or capsules. A syrup, elixir, or the likecan be used when a sweetened vehicle is employed. When indicated,suppositories or enema formulations may be the recommended route ofadministration.

Sustained or directed release compositions can be formulated, e.g.,liposomes or those wherein the active compound is protected withdifferentially degradable coatings, e.g., by microencapsulation,multiple coatings, etc. It is also possible to freeze-dry the newcompounds and use the lyophilisates obtained, for example, for thepreparation of products for injection.

Thus, the present invention provides for use of the Lipid-conjugates invarious dosage forms suitable for aerosol, rectal, vaginal,conjunctival, intravenous, intra-arterial, and sublingual routes ofadministration.

It will be appreciated that the actual preferred amounts of activecompound in a specific case will vary according to the specific compoundbeing utilized, the particular compositions formulated, the mode ofapplication, and the particular situs and organism being treated.Dosages for a given host can be determined using conventionalconsiderations, e.g., by customary comparison of the differentialactivities of the subject compounds and of a known agent, e.g., by meansof an appropriate, conventional pharmacological protocol.

Without further elaboration, it is believed that one skilled in the artcan, using the is preceding description, utilize the present inventionto its fullest extent. The following preferred specific embodiments are,therefore, to be construed as merely illustrative, and not limitative ofthe remainder of the disclosure in any way whatsoever.

EXAMPLES

The main abbreviations used in the examples below are:

-   HA=hyaluronic acid-   HYPE=dipalmitoyl-phosphatidyl-ethanolamine (PE) conjugated to HA    (also referred to as HyPE, HyalPE)-   CSA=chondroitin sulfate A-   CSAPE=PE conjugated to CSA (also referred to as CsAPE, CsaPE)-   CMC=carboxymethyl cellulose-   CMPE=PE conjugated to CMC-   HEPPE=PE conjugated to heparin (also referred to as HepPE, HePPE)-   DEXPE=PE conjugated to dextran-   AsPE=PE conjugates to aspirin-   HemPE=PE conjugated to Polygeline (haemaccel)-   HyDMPE=dimyristoyl PE linked to HA    Examples demonstrating the utility of lipid-conjugates in preventing    and treating disease are presented in PCT/US05/06591 filed    2-Mar.-2005, U.S. application Ser. No. 10/989,606 filed 17-Nov.-2004    and U.S. application Ser. No. 10/989,607 filed 17-Nov.-2004, which    are incorporated herein by reference in their entirety.

Example 1 Obstructive Respiratory Disease

The Lipid-conjugates are effective in the treatment of obstructiverespiratory disease. This is demonstrated for asthma in the Experiments1-8 below. In asthma, the impeded airflow is due to airway obstructionwhich is the result of constriction and obstruction of luminal vesselsof the lungs. One widely-accepted experimental system to investigateairway constriction is to induce smooth muscle preparations, isolatedfrom airways, to contract in the absence and presence of the drug.Another widely-accepted test of anti-asthma drug action is to use liveanimals which have asthma. This disease is present in animals which havebeen sensitized to an antigen and which can be monitored forexacerbation and recovery from asthmatic breathing using a bodyplethysmography.

In Experiments 1.1-1.3, the muscle preparation (tracheal rings) wasisolated from rats and in Experiment 1.4-1.5 from guinea pigs. Musclecontraction is measured by attachment of the muscle to a pressuretransducer, which works much like a spring. Induction of contractionoccurs when asthmatogenic substances are administered such asendothelin-1 (ET) an acetylcholine (AcCh).

Experiment 1.1: Isolated rat tracheal rings (in a linear array) werebathed in Krebs-Hanselet buffer (pH=7.4), and linked to a tensiontransducer. ET-1 was added to a final concentration as indicated, andthe tracheal ring contraction was determined by the change in the forceapplied to the tension transducer (FIG. 1.1A). Subsequently, the highestET concentration was used in testing the Lipid-conjugates to inhibit thesmooth muscle contraction. In this experiment (FIG. 1.1B), rat trachearings were incubated with the Lipid-conjugate HyPE at the indicatedconcentration for 1 hr. ET-1 was then added to a final concentration of1 μM and the ring contraction was determined as in Experiment 1.1A. Eachdatum is mean±S.D. of four separate experiments (4 rats).

Experiment 1.2: Rat trachea rings were incubated with 3 μM HYPE orhyaluronic acid (HA) alone, for 1 hr. ET-1 was then added to a finalconcentration of 1 μM (empty bars) or 10 μM (full bars) and the trachealring contraction was determined as in Experiment 1.1 (FIG. 1.2).

Experiment 1.3: The same as Experiment 1.2, but the tracheal ringcontraction was induced by 10 μM Acetyl Choline (AcCh), as shown in FIG.1.3.

Experiment 1.4: Guinea pig tracheal rings (in a linear array), immersedin a ringer bath, were connected to an apparatus measuring the length ofthe ring chain. CMPE or HEPPE was added to the bath 1 h prior to thestimulation of contraction by either Crotalus atrox (type II) enzyme orendothelin-1 as indicated (Table 1.1).

TABLE 1.1 Inhibition of Tracheal Ring Contraction by CMPE and HEPPEStimulant Lipid-conjugate % inhibition Phospholipase (0.5 μ/ml) CMPE (10μM) 100 ± 0.3  (crotalus atrox type II) Histamine (20 μM) CMPE (10 μM)69 ± 0.1 Histamine (20 μM) HEPPE (15 μM)  56 ± 0.05 Endothelin-1 (100nM) CMPE (10 μM) 92 ± 1.1

Experiment 1.5: Guinea pig tracheal rings were incubated with or withoutCMPE for 30 minutes prior to stimulation. The medium was collected after30 minutes and PGE₂ and TXB₂ were determined by radioimmunoassay (Table1.2). (n.d.=below limit of detection.)

TABLE 1.2 Inhibition of Tracheal Tissue PGE₂ and TBX₂ Production by CMPEPGE₂ TXB₂ Stimulant CMPE (ng/ml) (ng/ml) Hitsamine (40 μM) — 5.1 5.6Histamine (40 μM) 10 μM n.d. 1.75

Experiments 1.6-1.8 demonstrate the ability of Lipid-conjugates to exerttheir pharmacological effect in live animals. The following procedureswere applied in these experiments:

Inbred Brown Norway male rats (4 weeks old) obtained from Harlan, USA,were used in this study. The Hebrew University Animal Welfare Committeeapproved all protocols.

Induction of asthma: Asthma was induced in rats by sensitization withovalbumin (OVA, Sigma-Rehovot, Israel) according to a previouslydescribed protocol (33): On day 0 rats received a single subcutaneousinjection of 1 mg OVA+aluminum-hydroxide (200 mg/ml in 0.9% NaCl)(Sigma-Rehovot, Israel) and an intraperitoneal injection of 1 mlcontaining 6×109 heat-killed Bordetella Pertussis bacteria (PasteurMarieux, France). Repeated bronchial allergen challenge was performedfrom day 14 every other day for 1 month by inhalation of OVA (1 mg/ml in0.9% Normal Saline) for 5 minutes each time in a 20 L box connected toan ultrasonic nebulizer (LS 230 System Villeneuve Sur Lot, France).

Treatments: Rats were divided into 4 treatment groups: 1. Nosensitization and no treatment, used as Naïve control. 2.Sensitization+challenge with OVA and no treatment, used as positivecontrol. 3. Sensitization+challenge with OVA and treatment withLipid-conjugate (HyPE), either by sub-cutaneous (SC) injection orinhalation, before every challenge (HyPE). 4 (in part of theexperiments)−sensitization+challenge with OVA and treatment with SCinjection of dexamethasone 300 μg before each challenge (OVA/Dx). TheOVA/OVA group received 1 ml saline before each challenge.

Two modes of treatments with HyPE were employed: 1. The rats received SCinjection of 1 ml saline containing 15 mg HyPE (to obtain about 1 mg/mlbody fluid=20 μM). 2. The rats, placed unrestrained in a 20 liter boxconnected to an ultrasonic nebulizer, inhaled HyPE as follows: 5 ml of 1mg/ml HyPE was aerosolized into the 20 L cage, thus diluting the HyPE to0.25 μg/ml aerosol The rat respiratory rate was 120 breath/min, with atidal volume of about 1 ml, thus reaching ventilation of 120 ml/minute.If all the inhaled HyPE was absorbed, in 5 min (inhaling 600 ml), themaximal HyPE absorbed was 150 μg

In mode 1, all groups (5 rats in each) were treated and challenged asdescribed above on day 14, 16, 18 and 20, and pulmonary function (Penh)was assessed on day 20 before and 5 min after challenge (EAR).

In mode 2, each group (10 rats in each) were treated and challenged fromday 14, every other day, until day 45. Pulmonary function (Penh) wasassessed on day 20 before and 5 min and 8 h after challenge,corresponding to early and late asthmatic reaction (EAR and LAR,respectively).

Assessment of broncho-constriction: Unrestrained conscious rats wereplaced in a whole-body plethysmograph (Buxco Electronics Inc., Troy,N.Y., USA) connected to a pneumotach (EMKA Technologies, Type 0000) atone end, and to a 10 ml bottle at the other end. The pneumotach wasconnected to a preamplifier (model MAX2270, Buxco Electronics). Analoguesignals from the amplifier were converted to a digital signal by an ADcard (LPM-16 National Instruments, Austin, Tex., USA).Broncho-constriction measures were expressed as the enhanced pause(Penh). Penh=(PEF/PIF)*((Te−Tr)/Tr), where PEF=Peak Expiratory Flow,PIF=Peak Inspiratory Flow, Te=Expiratory Time, Tr=Relaxation time=timeof the pressure decay to 36% of total box pressure during expiration.

Broncho-alveolar lavage (BAL): On day 45 the rats were sacrificed bybleeding is through the abdominal aorta under anaesthesia withintra-peritoneal injection of sodium pentobarbital (100 mg/kg). The ratswere tracheotomized and incannulated through the trachea.Bronco-alveolar lavage (BAL) was collected by repeated washing of thelungs with 5 ml saline to a total of 50 ml.

Assessment of airway pathology: Susequent to collection of BAL, lungswere removed and inflated with 4% buffered formaldehyde under pressureof 20 cm H2O. The lungs were sliced longitudinally and embedded inparaffin. Histological sections 3 μm thick were cut and stained withhematoxylin and eosin for assessments of interstitial and peri-bronchialinflammation and airway smooth muscle thickening. Other slides werestained with Tri-chrome for assessment of sub-epithelial fibrosis (basalmembrane) and with PAS for epithelial cell mucus metaplasia.

Histological morphometry of airway structural changes was performedusing a computer program “ImageJ” (NIH Bethesda USA) on 3 randomlyselected slides from each mouse. Quantification of peribronchialcellular infiltrate in airway tissue was achieved through counting thenumbers of these cells in the 50-μm region beneath the epithelium of theairway in hematoxylin and eosin stained sections. Cells were expressedas number per millimeter of airway basal lamina length, which wasmeasured by tracing the basal lamina in calibrated digital images (43).Morphometric analysis of ASM and the basal membrane mass as indices oftheir thickening were performed as previously described (44). Briefly,measurements of the airway were obtained by tracing the digitalizedimages of interest. The outlines of the airway structures weresubsequently measured. All airways were evaluated for the followingmorphometric dimensions: length of the airway basement membrane of theepithelium (Lbm) and area of the ASM in the eosin hematoxylin stainedslides and the blue stain of the basal membrane of the Tri-chromestained slides. ASM cells or the basal membrane thickening werenormalized to the square of the Lbm (in μm2) to correct for differencesin airway size. Only large (>2,000 μm Lbm) and medium size airways(1.000-2,000 μm Lbm) were selected as it was shown that the mostsignificant pathological changes occur in these airways.

Protein expression of sPLA2 in lung tissue: Proteins were identified inhomogenized lung tissue (100 μg protein) using standard Western blot. Aspecific polyclonal antibody against Anti-sPLA2 antibody (Santa Cruz)diluted 1:500 (v/v) in TBST buffer+0.1% BSA. The immune reaction wasdetected by enhanced chemiluminescence (ECL).

Cysteinyl Leukotriene (CysLT): CysLT levels were measured in BAL using akit for direct enzyme immunoassay (EIA), according to manufacturer'sinstructions (Amersham Pharmacia Biotech U.K). The specificity of thekit was 100% for LTC4, 100% for LTD4, and 70% for LTE4. Result range wasbetween 0 to 48 pg.

Cell culture—Cells were isolated from the BAL were suspended in DMEMmedium supplemented with 10% fetal calf serum (FCS) and plated in a96-well plate at 106 cells/well. The cells were incubated for 2 hours in37° C., then non-adherent cells were removed by washing with PBS. Theadherent cells were re-suspended in DMEM supplemented with 10% FCS at106 cells/well and incubated for 48 hours. The culture medium was thencollected and assayed for determination of biochemical markers.

Nitric Oxide (NO) production—NO production by the BAL culturedmacrophages was determined by measuring their level in the culturemedium using the photometric method of Griess et al. (45).

TNFα production: TNFα production by the BAL cultured macrophages wasdetermined in the culture medium using radio-immunoassay (RIA) kits[Amersham-Pharamcia, UK).

Statistical Analysis: All data are expressed as mean±SEM. One way ANOVAwas used to compare treatment groups. Pair-wise comparisons wereperformed by the Tukey-Kramer HSD test (p=0.05). Where necessary, datawere log transformed before analysis to stabilized variances. In allanalyses P<0.05 was considered statistically significant.

Statistics: Statistical analysis was performed using statisticalsoftware (GB-STAT, Dynamic Microsystem Silver Spring Md., USA. Analyzisof variance (ANOVA) was used to assess difference of the results of thetreatment groups. A Tukey test was used to compare between each one ofthe treatment groups. A value of p<0.05 was considered as a significantdifference.

Expeiment 1.6—demonstrates that SC-administration of Lipid conjugatesconsiderably ameliorate OVA-induced broncho-constriction (FIG. 1.4,bronchoconstriction was induced in OVA-sensitized rats by inhalation ofOVA, and expressed by the difference in Penh measured before and 5 minafter allergen challenge. Each datum is Mean±SEM for 10 rats.Statistical significance: a−P<0.01; b, c−P<0.05), reduced the expressionof secretory phospholiapse (FIG. 1.5, the figure depicts Western blotand corresponding densitometry of sPLA₂ in lung homogenates of rats withOVA-induced asthma, treated as indicated. In panel B, for each enzymethe density values were normalized to corresponding Naïve), andprevented the production of the broncho-constricting lipid mediatorscysteinyl leukotrienes (FIG. 1.6, broncho-alveolar lavage (BAL) wascollected upon sacrifice and CysLT levels were determined by EIA, asdescribed in Methods. Each datum is Mean±SEM for 10 rats. Statisticalsignificance: a, b−P<0.01. No significant difference between HyPEtreated and the Naive rats).

Experiment 1.7 (aerosolic administration of HyPE) demonstrates thattreatment of the asthmatic rats by inhalation of the Lipid-conjugate,reduces protects the rats from sensitization by OVA, as it markedlyreduced OVA-induced broccho-constricyion in both the early and lateasthmatic reaction (FIG. 1.7, bronchoconstriction, expressed as thepercent change of Penh was induced in OVA-sensitized rats by inhalationof OVA, and measured before allergen challenge, 5 min and 8 h afterallergen challenge. Each datum is Mean±SEM for 10 rats. Two experimentswere performed for EAR. 5 rats were included in each group in the firstexperiment. The same experiment was repeated with 10 rats in each group,which were further used for determination of LAR. Combined statisticaltest for EAR yielded p<0.01 between Asthmatic and HyPE-treated; nosignificant difference between the HyPE-treated and the Naive orDx-treated groups. For LAR, p<0.01 between Asthmatic and HyPE-treated;no significant difference between the HyPE-treated and the Naive orDx-treated groups), inhibited the production of CysLT, potentbrocnho-constricting lipid mediator (FIG. 1.8, broncho-alveolar lavage(BAL) was collected upon sacrifice and CysLT levels were determined byEIA. Each datum is Mean±SEM for 10 rats. P<0.01 between asthmatic andHyPE-treated rats. No significant difference between HyPE treated andthe Naive rats), and of nitric oxide (NO), a characteristic constrictorof smooth muscle cells (FIG. 1.9, macrophages, collected from the BAL ofthe different groups, were cultured without further treatment with HyPEor Dx, and NO production was determined as described in Methods. Eachdatum is Mean±SEM for 10 rats. NO level was reduced compared toasthmatic and naïve rats by both HyPE, p<0.001 and p<0.001 respectivelyand by Dx p<0.001 and p<0.001, respectively.) These treatments alsoprevented the asthma-associated inflammation, as expressed by preventionof inflammatory cell infiltration and airway remodeling (FIG. 1.10, ratswere subjected to OVA inhalation every other day for 30 days. Fortreatment with HyPE, the rats inhaled HyPE aerosol for 5 min beforeevery allergen inhalation. The rats were sacrificed on Day 45.A—Staining with hematoxylin eosin for detection of inflammatory cellinfiltration and changes in smooth muscle cell (ASM) thickness.B—Staining of connective tissue (collagen) with Mason-Trichrom, fordetection of changes in basal membrane thickness. C—Staining withPeriodic Acid Schiff (PAS) for detection of mucus metaplasia ofrespiratory epithelial cells. 1, 2, 3 and 4 depict tissues of Naive,Asthmatic, HyPE-treated and Dx-treated rats, respectively, and FIG.1.11), and production of TNF-alfa by lung macrophages (FIG. 1.12,macrophages, collected from the BAL of the different groups, werecultured without further treatment with HyPE or Dx, and NO productionwas determined as described in Methods, Each datum is Mean±SEM for 10rats. p<0.001 between asthmatic and HyPE-treated rats. No significantdifference between HyPE-treated, Naive and Dx-treated rats).

Experiment 1.8, in which HyPE was given as aerosol to only beforechallenge to rats that had been sensitized by OVA (HyPE was not givenduring sensitization as in Experiment 1.7), demonstrates that inhalationof Lipid conjugates is effective in preventing allergen-inducedbroncho-condtriction in already asthmatic subjects when inhaled beforeallergen (OVA) challenge (FIG. 1.13, OVA-sensitized asthmatic ratsinhaled nebulized HyPE (1 mg/ml) for 5 minutes, or nebulized normalsaline. 30 minutes later all were challenged by inhalation of OVA (1mg/ml) for 5 minutes. Penh was measured before the treatments(baseline), and 5 minutes after each inhalation. Each datum is mean±SEMfor 5 rats. *,**, P<0.05), and revrese broncho-constricion (inducebroncho-dilation) when inhaled after allergen challege. FIG. 1.14:OVA-sensitized asthmatic rats challenged by imhalation of OVA (1 mg/ml)for 5 minutes. 30 minutes later they were treated by inhaltion ofnebulized HyPE inhalation (1 mg/ml) or nebulized or with normal salinefor 5 minutes. Penh was measured before challenge (baseline), and afterchallenge and treatment. Each datum is mean±SEM for 5 rats. *, P<0.05.

These experiments demonstrate that the Lipid-conjugates may be used forthe treatment of obstructive respiratory disease, alleviating airwaynarrowing by a plurality of mechanisms, including inhibition ofcontraction and reduction of airway obstructing infiltrates.

Example 2 Anti-Oxidant Therapy

The Lipid-conjugates are effective therapy for preventing oxidativedamage. This is demonstrated in Experiments 2.1-2.3. The noxious effectof peroxide free radicals on living tissue is known as oxidative damage.When cell membranes are the targets for this damaging process, membranedysfunction and instability result. Oxidative damage to blood proteins,particularly blood lipid proteins, results in their over-accumulation incells lining the vasculature, thus contributing to atherogenesis. Infact, oxidative cell damage is a major mechanism attributed to theprocess of aging or senescence.

Oxidative damage to proteins or cell membranes is commonly assessed byexposing these tissues to hydrogen peroxide produced by the enzymeglucose oxidase (GO), in the absence or presence of additional membranedestabilizing agents, such as PLA₂, or by exposure to divalent cations,such as copper.

Experiments 2.1-2.3 demonstrate the ability of Lipid-conjugates topreserve cells from oxidative damage, as judged by the cells' retentionof both arachidonic acid and of low molecular weight intracellularsubstances.

Experiment 2.1: Confluent BGM (green monkey kidney epithelial cells)were labeled with ³H-arachidonic acid, The cells were treated with CMPEfor 30 min prior to treatment with GO and PLA₂ (0.5 u/ml) (FIG. 2.1).

Experiment 2.2: BGM cells were labeled with ³⁵SO₄ overnight. The cellswere washed with DMEM (containing 10 mg/ml BSA) 4 times with PBS. Thecells were then incubated in DMEM supplemented with GO (an H₂O₂generation) for 90, and the culture medium was collected and counted for³⁵S radioactivity For treatment with CMPE cells were incubated withCMPE, at the indicated concentration for 30 min prior to introduction ofGO. Each datum is MEAN±SEM for 5 replications. *p<0.005; **p<0.001 (FIG.2.2).

Experiment 2.3: For demonstrating the ability of Lipid-conjugates toinhibit the oxidation of blood lipoprotein. LDL (0.1 μM) was incubatedin the absence and presence of various concentrations of HYPE or HA at37° C. At time zero 5 μM CuCl₂ was added to the dispersions and themixtures were continuously monitored for oxidation products at 245 nm(FIG. 2.3). The absorbance at 245 (OD units) is depicted as a functionof time (Schnitzer et al., Free Radical Biol Med 24; 1294-1303, 1998).

These experiments demonstrate that administration of Lipid-conjugates iseffective therapy in the prevention of tissue damage induced byoxidative stress (associated with free radical and hydrogen peroxideproduction) by a plurality of mechanisms, including inhibiting theoxidation of lipoprotein, as well as their uptake, inhibitingarachidonic acid release, and preserving the integrity of cell membranes(inhibiting GAG degradation), including red blood cell membranes.

Example 3 Lung Injury/Acute Respiratory Distress Syndrome (ARDS)

In acute respiratory distress syndrome (ARDS), which is usually inducedby bacterial endotoxins (LPS, LTA), a high production of injuriousmediators, particularly neutrophil-attracting chemokines, and cytokines,are produced by the lung microvascular endothelial cells (LMVEC). Todemonstrate the ability of the Lipid-conjugates to control theproduction of these injurious agents, LMVEC were treated with LPS(gram-positive bacterial endotoxin) and LTA (gram-negative bacterialendotoxin), in the absence and presence of Lipid-conjugates, and testedfor the subsequent production of cytokines and adhesion molecules,

To this end, human lung microvascular endothelial cells (LMVEC) werepurchased from CellSystems, Remagen, Germany at passage 4. The cellswere seeded in a density of 5000 cells^(−cm2) in T25 flasks andmaintained according to the manufacturer's specification in EGM-MV.Characterization of the LMVEC was performed on the basis of a positivestaining for uptake of acetylated LDL, Factor VIII related antigen andPECAM (CD31) expression as well as negative staining for alpha smoothmuscle actin. In each experiment the viability of LPS- andLTA-stimulated or HyPE-treated LMVEC was tested by trypan blueexclusion. The production and mRNA expression of cytokines and adhesionmolecules were determined as described in U.S. application Ser. No.10/989,606 filed 17-Nov.-2004, which is incorporated herein by referencein its entirety.

The production of the chemokines ENA-78, Gro-α and IL-8, secreted intothe culture medium of stimulated LMVEC, was measured by ELISAs accordingto the manufacturer's instructions.

For RNA isolation and Polymerase Chain Reaction by RT-PCR, confluentLMVEC were stimulated with medium as control or with LPS (1 μg^(−ml)) orLTA (10 μg^(−ml)) in the presence or absence of HyPE (10 μM). Total RNAwas isolated using Trizol-Reagent according to the manufacturer'sinstructions. Each RNA preparation was subjected to DNAse digestion toremove possible contaminations of genomic DNA. 1 μg of total RNA wasreverse transcribed using SuperScript™ II Preamplification Systemaccording to the manufacturer's instructions. Amplification of 0.5 μl ofcDNA was performed in a total volume of 25 μl containing 19.6 pmol ofeach chemokine primer, 5 mM of dNTPs, 2.5 U Taq Polymerase, 10 mM TrisHCl, 7.5 mM KCl, 1.5 mM MgCl₂. PCR reactions were initiated at 94° C.for 3 min, followed by 30 cycles of amplification, each consisting of94° C. for 1 min, 58° C. for 1 min, 72° C. for 2 min. At the end of theamplification cycles die products were incubated for 10 min at 72° C.Control samples were constructed either by omitting cDNA synthesis orwithout addition of cDNA. PCR products were separated on a 1% agarosegel. Real-time PCR: 500 ng of total RNA of each sample was in additionreverse-transcribed into cDNA for Real-time PCR analysis using 1stStrand cDNA Synthesis Kit according to the manufacturer's instructions(Roche). cDNA was diluted in 20 μl DEPC-treated water. DNA standardswere generated by PCR amplification of gene products, purification andquantification by spectrophotometry. Real time PCR of cDNA specimens andDNA standards were performed in a total volume of 25 μl in the presenceof 2 μl Light cycler-FastStart DNA Master SYBR GreenI reaction mix, 0.5μM of gen-specific primers and 4 mM MgCl₂. Standard curves weregenerated for all chemokines. PCR efficiency was assessed from theslopes of the standard curves and was found to be between 90% and 100%.Concentration of chemokine cDNA was calculated by linear regressionanalysis of all standard curves and was corrected for an equalexpression of GAPDH. At least five reproducible experiments wereperformed.

Adhesion molecules ICAM-1 and p-selectin were determined byfluorescence-activated cell sorter (FACS); Confluent LMVEC werestimulated with medium as control or with LPS (1 μg^(−ml)) or LTA (10μg^(−ml)) in the presence or absence of HyPE (10 μM). Thereafter cellswere harvested by T/E, extensively washed and monoclonal antibodiesdirected against the endothelial adhesion molecules ICAM-1 andP-selectin in dilutions of 1:20 were added for 30 min at 4° C. Inaddition unstimulated or stimulated cells were harvested as describedand preincubated for 20 min with HyPE (10 μM) and monoclonal antibodiesagainst TLR4. Cells were washed and incubated with an anti-mouseF(ab′)2, FITC conjugated secondary antibody. After washing cells wereanalyzed by FACS-scan.

Expression of NFκB was determined by Electrophorese mobility shift assay(EMSA); Confluent LMVEC were preincubated overnight in basal mediumcontaining 0.01% BSA. Thereafter they were stimulated or not fordifferent time periods with LPS, IL-1 or TNF-α in the presence orabsence of HyPE, and respective nuclear extracts were prepared.Oligonucleotides containing a NFkB consensus sequence (5′-AGT TGA GGGGAC TTT CCC AGG C-3′) were labeled to a specific activity >5×107cpm^(−μg) DNA. NF-kB-binding was performed in 10 mM HEPES, (pH=7,5), 0.5mM EDTA, 70 mM KCl, 2 mM DTT, 2% glycerol, 0.025% NP-40, 4% Ficoll, 0.1M PMSF, 1 mg^(−ml) BSA and 0.1 μg^(−μl) poly di/dc in a total volume of20 μl Nuclear extracts (10 μg) were incubated for 30 minutes at roomtemperature in the presence of 1 ng labeled oligonucleotide. DNA-proteincomplexes were resolved on 5% non-denaturating polyacrylamide gelselectrophoresed in low ionic strength buffer and visualized byautoradiography. Specificity of shifted bands was demonstrated by addinga cold NFkB consensus sequence or by supershift using anti-p65antibodies.

Experiment 3.1 demonstrates that the Lipid-conjugates are effective insuppressing the endotoxin-induced production and RNA expression of thechemokines IL-8, ENA-78 and Gro-α and their mRNA expression as shown inFIGS. 3.1, 3.2 and 3.3.

Experiment 3.2 demonstrates that the Lipid-conjugates are effective insuppressing the expression of the adhesion molecules ICAM-1 andE-selectin (FIG. 3.4).

Experiment 3.3 demonstrates that Lipid-conjugates are effective insuppressing the expression of NFκB, the transcription factor that isenhanced in endotoxin-induced injurious states (FIG. 3.5).

These results further demonstrate the therapeutic capacity of theLipid-conjugates in the treatment of ARDS and lung injuries, as well asother disease that share common mechanisms, such as peritonitis, kidneyfailure, organ transplantation and the like.

Example 4 Toxicity Tests

Experiment 4: The following compounds were tested: HyPE, CMPE, CSAPE andHepPE. The compounds were injected IP at one dose of 1000, 500 or 200mg/1 g body weight. Toxicity was evaluated after one week, by mortality,body weight, hematocrit, blood count (red and white cells), and visualexamination of internal organs after sacrifice. These were compared tocontrol, untreated mice. Each dose was applied to a group of three mice.No significant change in the above criteria was induced by treatmentwith these compounds, except for the HepPE, which induced hemorrhage.

The non-toxicity of the Lipid conjugates is demonstrated in Table 4.1and Table 4.2, depicting the results obtained for HyPE in acute (4.1)and long-term (4.2) toxicity tests.

TABLE 4.1 Acute toxicity Dose of HyPE Body weight RBC WBC Hematocrit(mg/kg body weight) (g) × 10⁶ × 10³ %   0.0 21.9 ± 0.2 22.6 ± 0.3 10.7 ±0.4 9.3 ± 0.3 45.0 ± 0.5 (control) 250  22.1 ± 0.4 23.1 ± 0.6 11.4 ± 0.17.7 ± 0.2 43.3 ± 0.7 500  21.4 ± 0.3 22.3 ± 0.4 11.5 ± 0.3 8.1 ± 1.344.7 ± 2.3 1000   21.7 ± 0.2 22.1 ± 0.2 10.9 ± 0.4 7.4 ± 0.6 40.3 ± 0.7RBC = red blood cells. WBC = white blood cells. Each datum is mean ±SEM.

For long-term toxicity test of HyPE, a group of 6 mice received a doseof 100 mg HyPE/Kg body weight, injected IP 3 times a week for 30 weeks(total of 180 mg to a mouse of 20 g). Toxicity was evaluated as forTable 4.1. No mortality, and no significant change in the above criteriawas induced by this treatment, compared to normal untreated mice (seeTable 4.1), as depicted in Table 4.2.

TABLE 4.2 Results at week 30: Body weight RBC WBC Hematocrit (g) × 10⁶ ×10³ % Control (untreated) 39.5 ± 3.1 10.9 ± 0.8 9.3 ± 0.6 45.0 ± 0.8rats HyPE-injected rats 39.0 ± 2.7 11.7 ± 0.7 8.1 ± 15  43.4 ± 4.9

Example 5 Synthesis Procedures

The procedures below are examples for synthesis of specific variants ofthe lipid-conjugates, and can be modified according to the desirablecompositions (e.g., changing the molar ratio between thelipid/phospholipid and the GAG, or the GAG size).

I. HyPE=phosphatidyl-ethanolamine (PE)-linked hyaluronic acid.

-   A. Truncating hyaluronic acid (HA):    -   Dissolve 20 g of HA in 12 L water, add 200 mg FeSO₄.7H₂O        dissolved in 20 ml water, add 400 ml H₂O₂ (30%), stir for 1.5 h.        Filter through 30 kD Filtron, Lyophilize. Yield: 16 g truncated        HA.-   B. Conjugation with PE (adjusted for 1 g):-   Prepare:    -   1. 10 g HA dissolved in 500 ml MES buffer, 0.1 M, pH=6.5    -   2. 1.0 g PE dissolved in 500 ml t-BuOH with 100 ml H₂O.        Mix the two solutions, add 1 g HOBT and 10 g EDC. Sonicate the        mixture in an ultrasonic bath for 3 h. Remove access free PE        (and EDC and HOBT) by extraction into organic phase (by addition        of chloroform and methanol to obtain a ratio of C/M/H₂O:1/1/1).        Separate the aqueous phase by a separation funnel. Repeat this        step twice. For final cleaning from reagents, filter through a        Filtron membrane (30 kD), and lyophilize.

Yield: about 8 g.

II. CSAPE=PE-linked chondroitin sulfate A (CSA):

-   Prepare:    -   1. 10 g CSA dissolved in 1.2 L MES buffer, 0.1 M, pH=6.5    -   2. 1 g PE dissolved in 120 ml chloroform/methanol: 1/1. Add 15        ml of a detergent (DDAB).        Mix 1 with 2, while stirring, add 1 g HOBT and 10 g EDC,        continue stirring thoroughly for a day at least. Remove access        free PE (and EDC and HOBT) by extraction into organic phase (by        addition of chloroform and methanol to obtain a ratio of        Chloroform/MeOH/EtOH/H₂O:1/1/0.75/1). Separate the aqueous phase        by a separation funnel. Repeat this step twice. Filter through a        Filtron membrane (30 kD), and lyophilize. To remove DDAB traces,        dissolve 1 g of dry product in 100 ml water and 100 ml MeOH, and        clean by ion exchanger using IR120 resin.-   Dialyse (to remove MeOH) and lyophilize.-   Yield: about 8 g.

Unexpected results showed that the sonication applied in the HyPEsynthesis, is an better substitute for the detergent in mixing theaqueous and lipid phases. Using sonication techniques simplifies thesynthesis and improves the purification of the product.

It will be appreciated by persons skilled in the art that the presentinvention is not limited by what has been particularly shown anddescribed herein above and that numerous modifications, all of whichfall within the scope of the present invention, exist. Rather, the scopeof the invention is defined by the claims which follow:

1. A method of decreasing the number or frequency of relapse episodes orreducing the incidence of disease-related symptoms of asthma in asubject, comprising the step of administering to a subject a compoundrepresented by the structure of the general formula I:

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; X is aglycosaminoglycan; and 2 to 200; and n is a number 2 to 200; whereinsaid spacer comprises —CO-alkylene-NH—, —CO-alkylene-CO— or acombination thereof and said spacer is linked to a hydroxyl orcarboxylic acid of said glycosaminoglycan; and wherein if Y is nothingthe phosphatidylethanolamine is directly linked to glycosaminoglycanevia a carboxylic group.
 2. The method according to claim 1, wherein saidglycosaminoglycan is hyaluronic acid.
 3. The method according to claim1, wherein said glycosaminoglycan is heparin.
 4. A method of treating asubject suffering from allergic rhinitis, comprising the step ofadministering to a compound represented by the structure of the generalformula I:

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; and X isa glycosaminoglycan; and n is a number 2 to 200; wherein said spacercomprises —CO-alkylene-NH—, —CO-alkylene-CO— or a combination thereofand said spacer is linked to a hydroxyl or carboxylic acid of saidglycosaminoglycan; and wherein if Y is nothing thephosphatidylethanolamine is directly linked to glycosaminoglycane via acarboxylic group.
 5. The method according to claim 4, wherein saidglycosaminoglycan is hyaluronic acid.
 6. The method according to claim4, wherein said glycosaminoglycan is heparin.
 7. A method of decreasingthe number or frequency of relapse episodes or reducing the incidence ofdisease-related symptoms of allergic rhinitis in a subject, comprisingthe step of administering to a subject a compound of formula I:

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; and X isa glycosaminoglycan; and n is a number 2 to 200; wherein said spacercomprises —CO-alkylene-NH—, —CO-alkylene-CO— or a combination thereofand said spacer is linked to a hydroxyl or carboxylic acid of saidglycosaminoglycan; and wherein if Y is nothing thephosphatidylethanolamine is directly linked to glycosaminoglycane via acarboxylic group.
 8. The method according to claim 7, wherein saidglycosaminoglycan is hyaluronic acid.
 9. The method according to claim7, wherein said glycosaminoglycan is heparin.
 10. A method of treating asubject suffering from chronic obstructive pulmonary disease, comprisingthe step of administering to a subject a compound of formula I:

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; and X isa glycosaminoglycan; and n is a number 2 to 200; wherein said spacercomprises —CO-alkylene-NH—, —CO-alkylene-CO— or a combination thereofand said spacer is linked to a hydroxyl or carboxylic acid of saidglycosaminoglycan; and wherein if Y is nothing thephosphatidylethanolamine is directly linked to glycosaminoglycane via acarboxylic group.
 11. The method according to claim 10, wherein saidglycosaminoglycan is hyaluronic acid.
 12. The method according to claim10, wherein said glycosaminoglycan is heparin.
 13. A method ofdecreasing the number or frequency or reducing the incidence ofdisease-related symptoms of chronic obstructive pulmonary disease in asubject, comprising the step of administering to a subject a compoundrepresented by the structure of the general formula I:

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; and X isa glycosaminoglycan; and n is a number 2 to 200; wherein said spacercomprises —CO-alkylene-NH—, —CO-alkylene-CO— or a combination thereofand said spacer is linked to a hydroxyl or carboxylic acid of saidglycosaminoglycan; and wherein if Y is nothing thephosphatidylethanolamine is directly linked to glycosaminoglycane via acarboxylic group.
 14. The method according to claim 13, wherein saidglycosaminoglycan is hyaluronic acid.
 15. The method according to claim13, wherein said glycosaminoglycan is heparin.
 16. The method accordingto claim 1, wherein said glycosaminoglycan is chondroitin sulfate. 17.The method according to claim 16, wherein said chondroitin sulfate ischondroitin-6-sulfate or chondroitin-4-sulfate.
 18. The method accordingto claim 1, wherein said phosphatidylethanolamine is dimyrisotylphosphatidylethanolamine or dipalmitoyl phosphatidylethanolamine. 19.The method according to claim 4, wherein said glycosaminoglycan ischondroitin sulfate.
 20. The method according to claim 19, wherein saidchondroitin sulfate is chondroitin-6-sulfate or chondroitin-4-sulfate.21. The method according to claim 4, wherein saidphosphatidylethanolamine is dimyrisotyl phosphatidylethanolamine ordipalmitoyl phosphatidylethanolamine.
 22. The method according to claim7, wherein said glycosaminoglycan is chondroitin sulfate.
 23. The methodaccording to claim 22, wherein said chondroitin sulfate ischondroitin-6-sulfate or chondroitin-4-sulfate.
 24. The method accordingto claim 7, wherein said phosphatidylethanolamine is dimyrisotylphosphatidylethanolamine or dipalmitoyl phosphatidylethanolamine. 25.The method according to claim 10, wherein said glycosaminoglycan ischondroitin sulfate.
 26. The method according to claim 25, wherein saidchondroitin sulfate is chondroitin-6-sulfate or chondroitin-4-sulfate.27. The method according to claim 10, wherein saidphosphatidylethanolamine is dimyrisotyl phosphatidylethanolamine ordipalmitoyl phosphatidylethanolamine.
 28. The method according to claim13, wherein said glycosaminoglycan is chondroitin sulfate.
 29. Themethod according to claim 28, wherein said chondroitin sulfate ischondroitin-6-sulfate or chondroitin-4-sulfate.
 30. The method accordingto claim 13, wherein said phosphatidylethanolamine is dimyrisotylphosphatidylethanolamine or dipalmitoyl phosphatidylethanolamine.
 31. Amethod of delaying the onset of symptoms of asthma in a subject,comprising the step of administering to a subject a compound representedby the structure of the general formula I:

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; and X isa glycosaminoglycan; and n is a number 2 to 200; wherein said spacercomprises —CO-alkylene-NH—, —CO-alkylene-CO— or a combination thereofand said spacer is linked to a hydroxyl or carboxylic acid of saidglycosaminoglycan; and wherein if Y is nothing thephosphatidylethanolamine is directly linked to glycosaminoglycane via acarboxylic group.
 32. The method according to claim 31, wherein saidglycosaminoglycan is hyaluronic acid.
 33. The method according to claim31, wherein said glycosaminoglycan is heparin.
 34. A method of delayingthe onset of symptoms of allergic rhinitis in a subject, comprising thestep of administering to a subject a compound represented by thestructure of the general formula I:

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; and X isa glycosaminoglycan; and n is a number 2 to 200; wherein said spacercomprises —CO-alkylene-NH—, —CO-alkylene-CO— or a combination thereofand said spacer is linked to a hydroxyl or carboxylic acid of saidglycosaminoglycan; and wherein if Y is nothing thephosphatidylethanolamine is directly linked to glycosaminoglycane via acarboxylic group.
 35. The method according to claim 34, wherein saidglycosaminoglycan is hyaluronic acid.
 36. The method according to claim34, wherein said glycosaminoglycan is heparin.
 37. A method of delayingthe onset of symptoms of chronic obstructive pulmonary disease in asubject, comprising the step of administering to a subject a compoundrepresented by the structure of the general formula I:

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; and X isa glycosaminoglycan; and n is a number 2 to 200; wherein said spacercomprises —CO-alkylene-NH—, —CO-alkylene-CO— or a combination thereofand said spacer is linked to a hydroxyl or carboxylic acid of saidglycosaminoglycan; and wherein if Y is nothing thephosphatidylethanolamine is directly linked to glycosaminoglycane via acarboxylic group.
 38. The method according to claim 37, wherein saidglycosaminoglycan is hyaluronic acid.
 39. The method according to claim37, wherein said glycosaminoglycan is heparin.
 40. The method accordingto claim 31, wherein said glycosaminoglycan is chondroitin sulfate. 41.The method according to claim 40, wherein said chondroitin sulfate ischondroitin-6-sulfate or chondroitin-4-sulfate.
 42. The method accordingto claim 31, wherein said phosphatidylethanolamine is dimyrisotylphosphatidylethanolamine or dipalmitoyl phosphatidylethanolamine. 43.The method according to claim 34, wherein said glycosaminoglycan ischondroitin sulfate.
 44. The method according to claim 43, wherein saidchondroitin sulfate is chondroitin-6-sulfate or chondroitin-4-sulfate.45. The method according to claim 34, wherein saidphosphatidylethanolamine is dimyrisotyl phosphatidylethanolamine ordipalmitoyl phosphatidylethanolamine.
 46. The method according to claim37, wherein said glycosaminoglycan is chondroitin sulfate.
 47. Themethod according to claim 46, wherein said chondroitin sulfate ischondroitin-6-sulfate or chondroitin-4-sulfate.
 48. The method accordingto claim 37, wherein said phosphatidylethanolamine is dimyrisotylphosphatidylethanolamine or dipalmitoyl phosphatidylethanolamine.
 49. Amethod of treating a subject suffering from asthma, comprising the stepof administering to a subject a compound of formula I:

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; and X isa glycosaminoglycan; and n is a number 2 to 200; wherein said spacercomprises —CO-alkylene-NH—, —CO-alkylene-CO— or a combination thereofand said spacer is linked to a hydroxyl or carboxylic acid of saidglycosaminoglycan; and wherein if Y is nothing thephosphatidylethanolamine is directly linked to glycosaminoglycane via acarboxylic group.
 50. The method according to claim 49, wherein saidglycosaminoglycan is hyaluronic acid.
 51. The method according to claim49, wherein said glycosaminoglycan is heparin.
 52. The method accordingto claim 49, wherein said glycosaminoglycan is chondroitin sulfate. 53.The method according to claim 46, wherein said chondroitin sulfate ischondroitin-6-sulfate or chondroitin-4-sulfate.
 54. The method accordingto claim 49, wherein said phosphatidylethanolamine is dimyrisotylphosphatidylethanolamine or dipalmitoyl phosphatidylethanolamine.